Catalogue Waters 2011 - 2012

2011/2012

Waters Quality Parts, Chromatography Columns and Supplies Catalog

Dear Colleague, We are pleased to present the 2011 Waters Quality Parts, Chromatography Columns and Supplies Catalog. Our catalog contains todays leading separations technology products. With over 50 years of market leadership we remain committed to the development and manufacture of products that solve todays most challenging separations problems to enable our customers success. We are driven by our core mission to provide enabling technologies in products that set the industry standard for performance and reproducibility. Waters provides the industrys broadest and deepest product offering manufactured to industry benc hmark quality requirements by our world-class manufacturing organization and supported by our highly skilled sales and support teams. We invite you to visit our new Online Order Center at www waters.com/order. T he online ordering experience has been improved to make it easier and faster for you to obtain pricing and availability when buying products. T his year for the fist time our catalog is available in digital form. You may access and download it at www.waters.com/catalog. T his past year we launched several innovative, enabling products including our new CSH technology that is the basis for the XSelect brand of HPLC columns and the ACQUITY CSH brand of UPLC columns, the Ostro Sample Preparation plate for the removal of phospholipids in biological samples and the Viridis line of columns for supercritical fluid chromatography. To learn more about how we can help you achieve your objectives please telephone one of our 54 worldwide offices and interact with our highly trained account specialists. Michael J. Yelle Senior Director, Chemistry Commercial Operations Waters Corporation Milford, MA, U.S.A.

CONTENTS

How to Order Sample Preparation Sample Vials and Accessories Filtration ACQUITY UPLC Columns and Consumables Analytical ChromatographyCover Image:Oasis HLB, a true binary mixed-mode, neutral copolymer, has a unique set of physicochemical properties that delivers the benefits of simplicity, speed, and reproducibility in SPE methods development. Its combination of water wettability and strong reversed-phase retentionenables high-throughput sample preparation in 96-well plate formats and engenders confidence in analytical results.

7 10 62 80 86 106 177 196 198 267 282 328 336

Preparative Chromatography Supercritical Fluid Chromatography (SFC) Bioseparations and Analyses Size-Exclusion Chromatography GPC & GFC Supplies and Hardware Alphabetical Index Part Number Index

WhaTS NEW

ACQUITY UPLC CSH Columns

XSelect HPLC Columns

Page 91CSH Technology is the next evolution of UPLC and HPLC particle tec hnology. It affords outstanding batc h-to-batc h reproducibility, extended pH stability and seamless transferability between UPLC, analytical HPLC and Preparative HPLC system platforms. T he ACQUITY UPLC CSH column family provides separation scientists wit h a wider range of selectivities as well as industry-leading performance in simple, LC/MS-friendly mobile phases.

Page 118XSelect HPLC columns are the next evolutionary step in Waters LC column tec hnology. Based on Charged Surface Hybrid (CSH) Tec hnology, this HPLC column family offers the widest range of selectivity choices and superior peak shape improvements for basic compounds when operated in acidic, low-ionic strength mobile phases (e.g., 0.1% formic acid).

Ostro Sample Preparation Products

Waters Positive Pressure-96 Processor

Page 53Ostro sample preparation products provide a novel solution for the cleanup of phospholipids in plasma and serum. Requiring minimal to no method development and using a simple protocol, this technology can be quickly implemented in order to optimize your laboratorys workflow. Providing cleaner, more reproducible extracts t han competitive phospholipid removal devices or tec hniques, the Ostro 96-well plate allows for more sensitive analyses, increased sample throughput, and reduced instrument downtime.

Page 51T he Waters Positive Pressure-96 Processor offers state of the art operation for 96-well and 1 cc flangeless cartridge formats. Each of the 96 holes in the processor is restricted in order to maintain constant pressure, even if all the plate well positions are not filled.

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Whats NeW

Protein-Pak Hi Res Ion-Exchange (IEX) Columns

ACQUITY UPLC BEH200 SEC Columns

Page 223The Protein-Pak Hi Res IEX column family consists of three ion-exchange column chemistries each contained on monodisperse, non-porous particles for the analysis of intact biomolecules including monoclonal antibodies, recombinant proteins, DNA/RNA and vaccine components.

Page 217ACQUIT Y UPLC analytical size-exclusion c hromatography (SEC) columns for the characterization of proteins and their aggregates. By applying the power of UPLC technology, you can now obtain accurate, precise, high-throughput quantitative analysis of proteins suc h as therapeutic mAbs.

Viridis Supercritical Fluid Chromatography (SFC) Columns

Online Order Center and Digital Catalog

Page 196Combining state-of-the-art media manufacturing with the industryleading column technology, Viridis Supercritical Fluid Chromatography (SFC) columns bring a new level of reproducibility to the world of laboratory-scale purification.

Page 7Waters Online Order Center streamlines the order process by placing product information, availability, tec hnical specifications and price in one central location. Combine the Online Order Center with Waters digital Chromatography Columns & Supplies Catalog, and shopping with Waters has never been easier.

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LITERaTURE ShOWCaSE

NEW ACQUITY UPLC CSH and XSelect HPLC Columns Brochure Lit. Co de 720003495EN

ACQUITY UPLC Columns Brochure Lit. Code 720001140EN

ACQUITY UPLC Column Strategies Brochure Lit. Code 720003271EN

XBridge HPLC Columns Brochure Lit. Code 720001255EN

SunFire HPLC Columns Brochure Lit. Code 720000875EN

Atlantis Columns Brochure Lit. Code 720002861EN

Prep OBD Columns Brochure Lit. Code 720002336EN

NEW Viridis SFC Columns Brochure Lit. Code 720003326EN

Oasis Sample Extraction Products Brochure Lit. Code 720001692EN

Sample Vials and Accessories Brochure Lit. Code 720001818EN

To view and download our latest literature, simply visit www.waters.com and enter the literature code in the search field (top right)

Sorbent Selection Guide for SPE Wall Chart Lit. Code 720002007EN

Analytical Columns Wall Chart Lit. Code 720002241EN

Prep OBD Wall Chart Lit. Code 720002117EN

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LITERATURE SHOWCASE

NEW Ostro Sample Preparation Products, Lit. Code 720003695EN

Waters DisQuE Products for QuEChERS, Lit. Code 720003048EN

PoraPak RXN for Post Synthesis Cleanup Brochure Lit. Code 720002747EN

Sample Preparation Solutions Brochure Lit. Code 720000848EN

Bioseparations and Analyses Catalog Lit. Code 720002148EN

Environmental Analysis Catalog Lit. Code 720002639EN

Food Testing Solutions Brochure Lit. Code 720002544EN

Environmental Analysis Solutions Brochure Lit. Code 720002766EN

Oasis Sample Preparation Application Notebook Lit. Code 720000609EN

Environmental Chromatography Methods Guide Lit. Code 720002543EN

Waters Vials Selector www.waters.com/vialsselector

Waters Column eSelectivity Chart Version 3.0 www.waters.com/selectivitychart

Waters Preparative OBD Column Calculator www.waters.com/prepcalculator

Waters Carbohydrate Method Selection Tool www.waters.com/amidemethod

NEW Waters Plate Selector www.waters.com/plateselector

NEW Waters Filter Selectorwww.waters.com/filterselector

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LITERATURE SHOWCASEWaters is committed to education and training. Learn from the best! Our expanding series of easy-to-read, well-illustrated, high-quality primers, written by experts, introduce, inform, and explain the latest technologies in analytical science.The Quest for Ultra Performance in Liquid Chromatography: Origins of UPLC TechnologyFrom the dawn of LC to the present day, drawn almost entirely from original sources and first-person accounts, this text reviews the first century of LC, showing how early the concepts of ultra performance were recognized and how many decades it took to reduce them to practice. An extensive glossary is included. Paperback, 54 pages, ISBN 978-1-879732-05-6Quest for Ultra Performance in Liquid Chromatography 715002098

Beginners Guide to Liquid ChromatographyOffering an uncomplicated introduction to the technology of liquid chromatography [LC], with a focus on HPLC, this basic book uses clear language, colorful diagrams, and a full glossary to acquaint readers with basic concepts and terminology. T his primer is suitable for younger science students as well as professionals new to LC. Paperback, 52 pages, ISBN 978-1-879732-02-5Beginners Guide to Liquid Chromatography 715001531

Beginners Guide to UPLC (Ultra-Performance Liquid Chromatography)Success is assured once new, experienced, and potential UPLC users learn from this volume the why and the how of U PLC Tec hnology principles. Scientists will gain t he confidence to apply t his knowledge in ways t hat en hance analytical productivity, streamline workflow, and advance scientific progress within their organizations. Paperback, 52 pages, ISBN 978-1-879732-07-0Beginners Guide to UPLC 715002099

NEW Comprehensive Guide to HILICHydrophilic Interaction ChromatographyT his tec hnology primer is designed to provide the reader with the basic insight of how to be successful with hydrophilic interaction c hromatography by understanding how the tec hnique works, the parameters that impact retention and selectivity, as well as the practical considerations necessary to successfully implement HILIC within a c hromatographic strategy. Paperback, 72 pages, ISBN 978-1-879732-08-7NEW Comprehensive Guide to HILIC 715002531

The Mass Spectrometry PrimerA wide range of topics related to a broad spectrum of mass spectrometric techniques is covered in this volume. In it, many frequently asked questions about the principles and practice of MS are answered. An extensive glossary explains MS terminology, and the benefits of coupling MS with chromatography are amply described. Paperback, 80 pages, ISBN 978-1-879732-04-1The Mass Spectrometry Primer 715001940

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How to order Waters Online Order CenterAn Improved Online Shoping Experience At Waters.comWaters.com is a convenient interface for both purchasing/procurement departments and scientific users to researc h, compare and purc hase consumables and spare parts online. We have now updated the web site to include an Order Center as a centralized point for all aspects of online shopping as well as adding options for easier connection to Enterprise Resource Planning (ERP) systems, or Spend Management Software packages. Ordering Waters Consumables and Spare Parts is quick and easy.

www.waters.com/order

Quick OrderExcellent starting point if you know the part numbers that need to be ordered.

How to Buy Service Plans, Instruments, or SoftwareOptions available for pricing and quotations.

Ordering FAQsFrequently asked questions explain all the new changes to the Order Center. Start here.

Certificate of Analysis (COA)Request a Certificate of Analysis for consumable products.

MSDS ( Material Safety Data Sheet)Safety precautions information on various products.

Shopping CartOrder multiple products easily, check real-time availability, and apply your company discount.

Freight PolicyDelivery time options and freight charges in each country.

Offers & DiscountsLook through all offers and discount codes available before you buy.

Technical Support (iRequest)Online support for technical queries on instrument/software.

Graphical Parts LocatorExploded instrument diagrams help you locate specific spare parts quickly.

Order HistoryFind previous ordering information.

Wish ListAdd part numbers for future projects, store frequently bought part numbers for easy repeat buying.

Step 1

Step 2

Step 3

[ The Order Center Process ]

Review product pricing Review product availability Review whether product requires hazardous shipping conditions Create a Wish List if you just want to email your choices

Register to buy online

Buy online Add discount coupons if available Automatically applies your company discount at checkout Choose shipping method Choose credit card payment (if available)

[ Benefits of e-Commerce ]Lower transaction costs Shorter order cycle time High Compliance levels Ease of finding products Account specific pricing Reduction of paper transactions Stock availability in real time Multiple Payment options Streamline internal process e-Order /e-Invoice

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How to orderWaters e-CommercePurchasing professionals regularly analyze their ordering and supply chains, seeking improved efficiencies, reduced costs and/or increased profitability achievable through more efficient procurement processes. If you are already using a Spend Management Solution or Internal ERP systems, you can leverage your investment by automating Waters product purc hases. Waters supports: 1. Catalogs via Punc hOut with many spend management providers. PunchOut can be connected directly with internal purchasing software, supporting many file formats like EDI (X12), cXML, xCBL (XML), etc 2. B2B e-procurement integration with spend management systems suc h as Ariba, SciQuest, SAP, Oracle, Perfect Commerce, Point Systems, ReQlogic, Direct Commerce, OB10, etc. 3. Access to product information with catalogs hosted in your IT environment using various business automation solutions. 4. We offer communication capabilities suc h as EDI, AS2, HTT P, HTT Ps, FT P, etc. We utilize Seeburger as the middleware to support electronic communications for B2C connections. 5. Posting electronic catalogs, suc h as Punc hOut (real-time pricing, availability, product pictures and specifications) or CIF (Catalog Interc hange Format), TXT (Text Format), XLS (Excel Format). Electronic catalogs can be provided in a timely manner. Please allow up to 90 days for implementation of electronic solutions (e-Orders / e-Invoices). To request information on integration with procurement solutions or for an electronic catalog, please contact Nancy Resteghini email: [email protected]

Waters Shopping FAQsW hether researc hing product information or making online purc hases, shopping on the Waters website has never been easier. T he Waters Chromatography Columns, Supplies and Parts catalogs are integrated into the product pages on the site, where you can see a list of available catalog items including current list price, and product number. Let Waters continue to be your online resource for the best chromatography columns, sample preparation products, accessories, and Waters Quality Parts. For your shopping convenience, please find our Online Shopping FAQs below.

How do I find instrument spare parts when I dont know what I am looking for?T here are two ways to find instrument spare parts: 1. Using the Graphical Parts Locator: a. b. c. Click the Services & Support tab > Find Spare Parts > and Graphical Parts Locator. Select a product from one of the dropdown lists, and select Waters Quality Parts Locator. Move your cursor over the picture of the instrument to discover orderable parts, or select a part from the displayed list. Continue to explore and drill down through the photos and graphics until you find the part you need. Click the part description, fill in a Quantity and click Add to Cart. Click the Services & Support tab > Find Spare Parts > and Spare Parts Catalog. Select a Value from one of the Menus (Product Category etc.) to begin. Continue to select values from the various menus to refine the results until you locate the desired part. Fill in a Quantity and click Add to Cart.

How do I get started? You may search for, see local list pricing, and add products to your shopping cart without login. Login is now required when you are ready to check out. If you have not logged in but added products to your cart, you will be prompted to login when you proceed to check out. If you have previously registered and have a login ID, you may log in at any time. If you have never registered; Click Sign-Up on the Login screen and complete our registration form. Registering allows ongoing access to Waters events, information, shopping, and support.

2. Using the Spare Parts Catalog: a. b. c.

How do I find chemistry related consumable products when I dont know what I am looking for?T here are two ways to find consumable products: a. Click on the Products tab (upper left corner) and select our Consumables & Columns catalog to look for specific products & prices in our catalog. Once you have refined your search and found the product you are looking for simply fill in a Quantity and click Add to Cart Click in the Global Search field, type in a search term ex; ACQUITY UPLC or a product part number and choose a pre-selected Facet Search> Columns & Consumables Columns > Select a value from the new filter bar design (Chemistry, Particle Size etc.) displaying relevant facets and value selection that can be used to drill down to your desired result. b. Continue to select values from the various menus to refine the results until you locate the desired product, click on the product header (for more details) or simply fill in a Quantity and click Add to Cart.

I already know my part number1. Click Cart in the upper right corner of the website. 2. Using the Add to Cart box: enter the Product Number, Quantity and click Add to Cart. Your items are added directly to the shopping cart.

b.

How do I use Wish List(s)?You can add frequently ordered products to a Wish List to expedite ordering on future orders. To manage (add/modify/delete) Wish List(s) click on My Account located in the upper right menu bar and select Wish List from the options list on the left hand side of the page. From here you may create, delete, and rename multiple Wish Lists To add a product to a Wish List:

a.

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hOW TO ORdER

1. Click Cart to display your shopping cart. 2. Place a c heck mark next to the Product Name and click Add to Wish List. If you have created multiple lists, select the list you would like to add the product to and click Add to Wish List. To remove an item in a Wish List: 1. Click My Account located in the upper right menu bar and select Wish List from the options list on the left hand side of the page and select Wish List. Place a check mark next to the Product Name and click Remove. You may also copy the selected item to other lists by clicking on Copy to List and selecting the list you would like to copy the selected product to. To order using a Wish List: 1. Click My Account located in the upper right menu bar and select Wish List from the options list on the left hand side of the page. If you have multiple shopping lists, choose a list from the View List box. 2. Place a check mark in the box next to the Product Name and click Add to Cart. A message displays: The selected item(s) have been successfully added to your cart. 3. Click Return to Wish List. Click the Close button then Cart to return to your shopping cart.

W hat if I am unable to purchase online or have a purchase approval process?Waters has provided the ability to email a copy of the Cart contents to someone else with your comments using our Share Cart feature. Sending this information is especially helpful for those who do not have the ability to directly purchase online. You can inform your purchasing department, local distributor or Waters Representative of your interest or needs. It should not be considered an ordering process but may help you to expedite your order. To use this feature: 1. Click Cart. 2. Click Share Cart; enter the email address and a brief message if desired. 3. Click Send.

How do I see related items?1. Click the Products tab > Chromatography Consumables & Columns > then select a Product Area. 2. Select a product grouping to refine your results and then select the Products tab. 3. W hen you find your Product, click the Product description link and if there is a related product, a related products section is displayed at the top of the page. NOTE: Not all products have related products available.

How do I check out?1. Click on the Cart icon when you have finished shopping 2. Review cart content, add web promotion discount codes in the Discounts box, and click Proceed to Checkout. 3. Step 1 - Shipping, Billing, and Payment Select a shipping address from your stored address list (if applicable). If a new address is required and you are a registered user, select the Enter a new address button, and complete the new address form*. Next, select your preferred shipping method,** add attention line and/or special instruction information if appropriate. T hen verify your billing address information following the same step noted above for a new address. Next, select a payment choice and complete the required payment method fields. Click Continue Checkout.* New addresses may take up to 24 hours for account information verification and processing. During this time you may still submit an order for processing. ** Products containing hazardous materials may be shipped separately and incur additional shipping costs.

How do I view my order history?To access your order history, from the Waters Home Page (after login): 1. Click My Account, and then click Order History. 2. Find your orders by indicating one of the following: Date range (from/to), Waters Order Number or your Purchase Order Number, then click Search. 3. Click on the Order Number link to display specific order detail.

I would like to order an entire system/instrument/ software/Service Plans but dont see these options.At this time such Waters does not offer ordering of these products via our online shopping center. Please follow this link (Plans/Instruments/Software link here) to request pricing and a local Waters sales representative will contact you shortly.

4. Step 2 Order Review Review your order detail for completeness. Discounts discount amounts appearing in the discount column are representative of web promotion codes you may have entered while viewing your shopping cart. (T hese discounts may also be added to your order here in Step 2). Invalid promotion codes may be the result of ineligibility, or an expired promotion. Other Discounts If your company has a contractual discount with Waters or your order is eligible for other promotions, these discounts may already be automatically applied to your order during Step 2 (Order Review) of our Check out process. T he best discount available will always be applied to your order. 5. Step 3 Order Confirmation You may print a copy of your order confirmation by clicking on the Print button once an order number has been created. An email confirmation will automatically be sent to the email address provided during the check out process. 6. Accept Waters Standard Terms & Conditions by selecting the Terms & Conditions I agree box and click the Place Order button.

Common Terminology & DefinitionsNot Available online T his part is not available for direct sale to customers. Please contact your Waters Local Representative for more information. Orderable Part This part is available and may be ordered through our Waters Online Order Center. Obsolete Part with no Replacement An obsolete part is no longer manufactured and may not be ordered from Waters. Also, there is no replacement for this part. Please contact your Waters Local Representative for further detail. Discontinued Part with Replacement T his indicates the part number you should order in place of an obsolete or discontinued part number. T he original part number has c hanged but the replacement part may be used in the same application.

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[ Sample Preparation ]The Importance of Sample Preparation ..................................................................................................................................................... 12 Sorbent Selection Guide ............................................................................................................................................................................ 13 Oasis Solid-Phase Extraction (SPE) Products Introduction ................................................................................................................................................................................. 16 Oasis MCX for Basic Compounds ............................................................................................................................................... 18 Oasis MAX for Acidic Compounds ............................................................................................................................................. 19 Oasis WCX for Strong Basic Compounds................................................................................................................................... 21 Oasis WAX for Strong Acidic Compounds ................................................................................................................................ 22 Oasis HLB for Reversed-Phase SPE ............................................................................................................................................ 23 Oasis Elution Plates.................................................................................................................................................................. 26 Oasis 96-Well High-Throughput Extraction Plates ................................................................................................................... 28 Sensitivity and Precision ........................................................................................................................................................... 30 Sorbent Amount and Solvent Selection for the Generic Method ............................................................................................................................................................... 31 Automation ................................................................................................................................................................................. 31 Oasis On-Line Columns .............................................................................................................................................................. 32 Oasis Glass Cartridges for PPT Detection Levels ...................................................................................................................... 33 Oasis Symbiosis On-Line Cartridges ......................................................................................................................................... 33 Oasis Product Selection Guide ................................................................................................................................................... 34 Oasis Sorbent Selection Tools ................................................................................................................................................... 35 Ordering Information ................................................................................................................................................................. 36 NEW Waters Positive Pressure -96 Processor ........................................................................................................................... 38 Sep-Pak Solid-Phase Extraction (SPE) Products Introduction ................................................................................................................................................................................. 39 Sep-Pak Sorbent Selection Guide .............................................................................................................................................. 40 Sep-Pak Cartridge Selection Guide ............................................................................................................................................ 42 Sep-Pak 96-well Plates .............................................................................................................................................................. 42 Sep-Pak 96-well Elution Plate................................................................................................................................................. 43 Sep-Pak Specialty Chemistries ................................................................................................................................................... 44 Sep-Pak DNPH-Silica Cartridges......................................................................................................................................... 45 Ozone Scrubber Cartridges ................................................................................................................................................ 46 Waters XPoSure Cartridges ................................................................................................................................................ 46 PoraPak RDX Cartridges ...................................................................................................................................................... 47 Sep-Pak Dry SPE Cartridges................................................................................................................................................ 47 Certified Sep-Pak Cartridges ...................................................................................................................................................... 48 Certified Sep-Pak Sorbent Selection Guide ............................................................................................................................... 49 Waters Extraction Manifold ........................................................................................................................................................ 50 NEW Waters Positive Pressure -96 Processor ........................................................................................................................... 51 Environmental and Food Solution Kits ..................................................................................................................................................... 52 NEW Ostro Sample Preparation Products.................................................................................................................................................. 53 DisQuE Dispersive Sample Preparation Kit ............................................................................................................................................... 54 PoraPak RXN Cartridges for Post Synthesis Cleanup................................................................................................................................ 56

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[ Sample Vials and Accessories ]Waters Autosampler Vials ......................................................................................................................................................................... 62 Choosing the Right Vial and Septum for Your Application ..................................................................................................................... 62 Vial Closures Guide .................................................................................................................................................................................... 63 Vial Selection Guide................................................................................................................................................................................... 63 Screw Cap 12 x 32 mm Vials for ACQUITY UPLC Systems .................................................................................................................... 64 Waters ACQUITY UPLC System Sample Manager .................................................................................................................................... 65 Plates for ACQUITY UPLC Systems ........................................................................................................................................................... 66 Glass Inserts for 96-Well Plates ............................................................................................................................................................... 66 Heat Seals................................................................................................................................................................................................... 67 Most Commonly Used Vials for Alliance Systems .................................................................................................................................. 68 Plates for Alliance HT Systems................................................................................................................................................................. 69 Screw Cap 12 x 32 mm Vials for Alliance Systems ............................................................................................................................... 70 Snap Cap 12 x 32 mm Vials for Alliance Systems................................................................................................................................. 71 Vials for Waters 717 Autosampler........................................................................................................................................................... 72 Vials for Waters 2707 Autosampler ........................................................................................................................................................ 73 Screw Cap 12 x 32 mm Vials for Compatible Systems .......................................................................................................................... 74 Snap and Crimp Cap 12 x 32 mm (9 mm Cap) Vials for Compatible Systems..................................................................................... 75 15 x 45 mm Vials for Compatible Systems ............................................................................................................................................ 76 GPC 2000 Vials.......................................................................................................................................................................................... 76 Aqua Analyis System Vials ....................................................................................................................................................................... 76 Settings for Alliance 2690 and 2695 Vials and Low Volume Inserts (LVI) ......................................................................................... 77 Vial Descriptions ........................................................................................................................................................................................ 78 Vials Troubleshooting Guide ...................................................................................................................................................................... 79

[ Filtration ]Sirocco Protein Precipitation Plate............................................................................................................................................................ 80 Pall Life Sciences Sample and Solvent Filtration Products ..................................................................................................................... 81 Filter Design and Membrane Choices........................................................................................................................................................ 82 Part Number Cross Reference .................................................................................................................................................................... 83 Solvent Filtration Membranes ................................................................................................................................................................... 83 Solvent Filtration Apparatus ..................................................................................................................................................................... 83 Syringe Filters Product List ....................................................................................................................................................................... 84

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SaMPLE PREPaRaTION

T he Importance of Sample Preparation

In the last two decades, dramatic advances in analytical instrumentation and laboratory information management systems shifted the analysts predominant tasks from assay measurements to sample preparation and data processing. As the stringency of requirements for higher sensitivity, selectivity, accuracy, precision, and number of samples to be processed has escalated, the corresponding increases in speed and sophistication of analysis and data collection have outpaced improvements in the many traditional techniques of sample collection and preparation. By some estimates, 75 to 80% of the work activity and operating cost in a contemporary analytical lab is spent processing and preparing samples for introduction or injection into an analytical separation and/or measurement device. Clearly, efforts directed and products designed to streamline sample preparation protocols are essential to future progress in analytical science.

Benefits of Solid-Phase Extraction (SPE) CartridgesW hen compared to other sample preparation processes, solid-phase extraction using SPE cartridges offers:

lower cost lower solvent consumption greater recoveries lower reagent consumption minimal sample transfer less apparatus faster protocol fewer steps greater safety less exposure to toxic agents greater accuracy no cross contamination

no emulsion problems less sample handling fewer steps no transporting of samples to lab direct field sampling reduced harm to labile samples minimal evaporation minimal glass breakage less glassware used, less to wash

Goals of Sample PreparationSuccessful sample preparation for most analytical techniques [HPLC, GC, spectrophotometry, RIA, etc.] has a threefold objective: namely, to provide the sample component of interest

in solution free from interfering matrix elements at a concentration appropriate for detection or measurement.

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Sample Preparation

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SaMPLE PREPaRaTION

SPE Application Area/Sorbent Selection GuideMatrix Key

Air

Petroleum

Biological

Soil

Water

Food

Sorbent/Surface

Common Sample Matrices

Description

Typical Applications

Properties

Reversed-PhaseOasis HLBN-VinylpyrrolidoneDVB copolymer Strongly hydrophobic, water-wettable polymer with unique hydrophilic-lipophilic balance. Maintains high retention and capacity even if it runs dry after conditioning. Stable in organic solvents. Strongly hydrophobic, silica-based bonded phase used to adsorb analytes from aqueous solutions. Trifunctional bonding chemistry for increased hydrolytic stability. Drugsandmetabolitesinbiofluids Isolationofpeptidesandoligonucleotides High-throughputbiopolymerdesalting Traceorganics,prioritypollutants,endocrinedisrupters JPMHLW official food methods: antibiotics, pesticides JPMHLW official methods for pesticides in water JPMHLW official methods for odorants in water Particle sizes: 30 and 60 m Pore size: 80 Surface area: 830 m2/g pH range: 014 Particle size: 3755 m Pore size: 125 Surface area: 325 m2/g Carbon load: 17% pH range: 28 Particle size: 55105 m Pore size: 125 Surface area: 325 m2/g Carbon load: 12% pH range: 28 Particle size: 3755 m Pore size: 125 Surface area: 325 m2/g Carbon load: 9% pH range: 28 Particle size: 3755 m Pore size: 125 Surface area: 325 m2/g Carbon load: 2.7% pH range: 28

Sep-Pak tC18Silica SiC18H37

Sep-Pak C18Silica Si(CH3)2C18H37

Hydrophobic, silica-based bonded phase used to adsorb analytes from aqueous solutions. Monofunctional bonding provides alternate selectivity versus tC18.

Lipidfractionation;gangliosideisolation Organicacidsinfruitjuice,wine JPMHLW and CDFA official methods: pesticides in food Naturalproducts AOAC methods for food colors, sugars Drugsandtheirmetabolitesinbiofluids Peptidesinserumandplasma

Sep-Pak C8Silica Si(CH3)2C8H17

Moderately hydrophobic, silica-based bonded phase used in methods when less retention than that of HLB or C18 is required.

Sep-Pak tC2Silica SiC2H5

Weakly hydrophobic, silica-based bonded phase used in methods when less retention than that of C8 is required. Trifunctional bonding chemistry for increased hydrolytic stability.

ApplicationsaresimilartothoseofC18 and C8

Reversed- or Normal-PhaseSep-Pak AminopropylSilica Si(CH2)3NH2 Moderately polar, silica-based bonded phase with weakly basic surface. Can be used as a polar sorbent with different selectivity for acidic/basic analytes or as a weak anion exc hanges in aqueous medium below pH 8. Silica-based bonded phase with low hydrophobicity can be used as a less polar alternative to silica or as a less hydrophobic alternative to C18 or C8. Phenols,phenolicpigments,naturalproducts Petroleumfractionation Saccharides Drugsanddrugmetabolites JPMHLW official methods for pesticides in food Drugsandtheirmetabolites Pesticides Particle size: 55105 m Pore size: 125 Surface area: 325 m2/g Carbon load: 3.5% pH range: 28 Particle size: 55105 m Pore size: 125 Surface area: 325 m2/g Carbon load: 6.5% pH range: 28 Particle size: 3755 m Pore size: 300 Surface area: 100 m2/g Carbon load: 2% pH range: 28

Sep-Pak CyanopropylSilica Si(CH3)(CH2)3CN

Sep-Pak DiolSilica Si(CH2)3OCH2CH(OH) CH2OH

Moderately polar, neutral, silica-based bonded phase. Used in normal-phase applications where acidic c haracter of silica is undesirable or as a weakly hydrophobic phase in aqueous media.

Antibioticsincosmetics ProteinandpeptideisolationbyHIC [ hydrophobic-interaction chromatography]

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Sample Preparation

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SaMPLE PREPaRaTIONCommon Sample Matrices

Sorbent/Surface

Description


Properties

Normal-PhaseSep-Pak SilicaSiO2 Polar sorbent binds analytes in non-aqueous solvents. Also used as an intermediate-strength cation exchanges in aqueous media and as a support for liquid-liquid partition separations. Polar, highly active, weakly basic sorbent for the adsorption of low to moderately polar species from non-aqueous solutions. Vitaminsandfoodadditives Lipidclassification Syntheticorganiccompounds Naturalproducts,plantpigments JPMHLW official methods for pesticides in food AOAC and EPA official methods for pesticides JPMHLW official methods for pesticides in food Polychlorinatedbiphenyls[ PCBs]intransformeroil Particle size: 55105 m Pore size: 125 Surface area: 325 m2/g Activity: High [ 3.2% water] Particle size: 50200 m Pore size: 60 Activity: High [ 2.5% water] pH of 10% aqueous slurry: 8.5 Particle size: 50300 m Pore size: 120 Activity: High, 1 on Brockmann scale [ 1.5% water] pH of 10% aqueous slurry: A: 4 N: 7.5 B: 10

Sep-Pak FlorisilMgOSiO2

Sep-Pak Alumina (A,N,B)Al2O3

Highly surface-active polar, acidic [A], neutral [N], and basic [B] sorbents. Exhibits specific pi-electron interactions with aromatic hydrocarbons. Acidic and basic alumina are also low-capacity ion exchangers in aqueous media, unaffected by high-energy radioactivity.

Petroleum,syntheticcrudeoilfractionation[N] Radioactivecompoundisolation,isotopegenerators[A,B] Phospholipids,steroids,catecholamines[B] Food,feedadditives[A,N],syntheticorganiccompounds[N] Pesticide,herbicide,prioritypollutantisolation[N,B] Alternative to official AOAC and EPA methods [A,N,B]

DispersiveDisQuESpecific Application Prepackaged centrifuge tube containing anhydrous sodium acetate/anhydrous magnesium sulphate and anhydrous magnesium sulphate/primaryl secondary amine sorbent. Used for dispersive solid-phase extraction, commonly referred to as QuEChERS. FoodSafety:Monitorcommoditiesforchemicalresidues to ensure compliance with established regulation FoodQualityandManufacture:Qualitycontrolof manufactured products AOACMethod2007.01 Properties: DisQuE Extraction [Tube 1]: A 50 mL centrifuge tube containing 1.5 g anhydrous sodium acetate and 6 g of anhydrous magnesium sulphate. DisQuE Clean-Up [Tube 2]: A 2 mL centrifuge tube containing150 mg anhydrous magnesium sulphate and 50 mg of primary secondary amine [PSA] sorbent.

Ion-Exchange and Mixed-ModeOasis MCXN-Vinylpyrrolidone DVB copolymer SO3H Mixed-mode, reversed-phase/strong cation-exchange, water-wettable polymer. Highly selective for bases. Stable in organic solvents. Basicdrugsfrombiofluidsandtissueextracts Drugmonitoring:screening,identification,confirmation, and quantitation Pesticides,herbicides Stronglybasiccompoundsinbiofluidsandtissueextracts Drugmonitoring:screening,identification,confirmation, and quantitation JPMHLW official method for streptomycin and dihydrostreptomycin in vegetable crops Isolationofcationicproteins Pesticides,herbicides Steroids Inorganiccationsinenvironmentalsamples Particle sizes: 30 and 60 m Pore size: 80 Surface area: 830 m2/g pH range: 014 [pKa: < 1] IEX capacity: 1 meq/g Particle sizes: 30 and 60 m Pore size: 80 Surface area: 830 m2/g pH range: 014 [pKa: ~ 5] IEX capacity: 0.75 meq/g Particle sizes: 3755 m Pore size: 300 pH range: 29 Ligand density: 350 moles/g Protein-binding capacity: 175 mg Cytochrome c/g Particle sizes: 30 and 60 m Pore size: 80 Surface area: 830 m2/g pH range: 014 [pKa: > 18] IEX capacity: 0.25 meq/g Particle sizes: 3755 m Pore size: 300 pH range: 29 Ligand density: 220 moles/g Protein-binding capacity: 200 mg BSA/g Particle sizes: 30 and 60 m Pore size: 80 Surface area: 830 m2/g pH range: 014 [pKa: ~ 6] IEX capacity: 0.6 meq/g Particle sizes: 37-55 m Pore size: 60 Surface area: 450 m2/g pH range: 2-9 IEX capacity: 1.75 meg/g

Oasis WCXN-Vinylpyrrolidone DVB copolymer COOH

Mixed-mode, reversed-phase/weak cation-exchange, water-wettable polymer. Used to retain and release strong bases. Stable in organic solvents.

Sep-Pak Accell Plus CMAcrylic acid/acrylamide copolymer on diol-silica COO Na+

Silica-based, hydrophilic, weak cation exchanger with large pore size used to extract cationic analytes in aqueous and non-aqueous solutions.

Oasis MAXN-Vinylpyrrolidone DVB copolymer CH2N(CH3)2C4H9+

Mixed-mode, reversed-phase/strong anion-exchange, water-wettable polymer. Selective for acids. Stable in organic solvents.

Acidiccompoundsandmetabolitesinbiofluidsand tissue extracts Drugmonitoring:screening,identification,confirmation, and quantitation Foodadditivesandcontaminants Isolationofanionicproteins Acidicpigmentsinwine,fruitjuices,foodextracts Phenoliccompounds Peptidepoolfractionation Inorganicanionsinenvironmentalsamples

Sep-Pak Accell Plus QMAAcrylamideacrylic acid copolymer bound to diol-silicaC(O) NH(CH2)3N(CH3)3+ Cl-

Silica-based, hydrophilic, strong anion exchanger with large pore size used to extract anionic analytes in aqueous and non-aqueous solutions.

Oasis WAXN-VinylpyrrolidoneDVB copolymerCH2piperazine

Mixed-mode, reversed-phase/weak anion-exchange, water wettable polymer. Used to retain and release strong acids. Stable in organic solvents.

Stronglyacidiccompoundsandmetabolitesinbiofluids and tissue extracts Drugmonitoring:screening,identification,confirmation, and quantitation Emergingcontaminants Strongaffinityforfattyacids,polarpigments,andsugars Potentialforadsorptionbychelation

Sep-Pak PSASiC2H4NHC2H4NH2

Silica-based bonded phase containing primary and secondary amines. Similar selectivity to aminopropyl but with higher pKas and increased ion-exchange capacity.

14

Sample Preparation

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SaMPLE PREPaRaTIONCommon Sample Matrices

Sorbent/Surface

Description


Properties

Products for Environmental and Food AnalysisSep-Pak DNPHDiphenylhydrazine coated on silica Acidified dinitrophenylhydrazine reagent coated on silica used for collection of air samples. Aldehydes and ketones react in situ to form hydrazone derivatives; these are then eluted and quantitated by HPLC analysis. Acidified dinitrophenylhydrazine reagent coated on silica used for collection of air samples. Aldehydes and ketones react in situ to form hydrazone derivatives; these are then eluted and quantitated by HPLC analysis. Larger particle size optimized for low-pressure personal air monitors. Potassium iodide cartridge is used in series with Sep-Pak DNPH and X PoSure Aldehyde Sampler cartridges to remove ozone interferences. High-capacity desiccant used to remove residual water from normal-phase SPE extracts [in water-immiscible organic solvents]. EPA Method TO11A; ASTM D5197 for carbonyl compounds in air JPMOE Official Methods for aldehydes: odor in out door air and in exhaust gas JPMHLW official methods for aldehydes in indoor air EPA Methods TO11A and IP6A, ASTM D5197 for carbonyl compounds in air NIOSH Method 2532 for glutaraldehyde in air EPA Method IP6A and ASTM D5197 for carbonyl compounds in air Generalpurpose Particle size: 55105 m Pore size: 125 Recommended maximum capacity: 75 g [2.5 moles] formaldehyde/cartridge Particle size: 5001000 m Pore size: 125 Recommended maximum capacity: 70 g [2.3 moles] formaldehyde/cartridge Quantity: 1.4 g KI Capacity: 4.2 mmoles ozone/cartridge [theoretical] Quantity: 2.85 g anhydrous Na2SO4 T heoretical capacity: 3.6 g H2O

Sep-Pak X PoSure Aldehyde SamplerDiphenylhydrazine coated on silica

Sep-Pak Ozone ScrubberPotassium iodide

Sep-Pak DryAnhydrous Sodium Sulfate

Sep-Pak Porapak RDXDVBN-Vinylpyrrolidone copolymer

Hydrophobic polymer for explosives analysis.

Explosivesingroundandsurfacewateratppblevel EPA Method 8330 for explosives

Particle size: 125150 m Pore size: 200 Typical sample volume: 500 mL water/cartridge Particle size: 80 m Quantity: 265 mg/cartridge

Sep-Pak PS2StyreneDVB copolymer

Very hydrophobic copolymer designed for multi-residue pesticide analysis in water samples.

JPMHLW official methods for pesticides in water JPMHLW official methods for pesticides in food

Sep-Pak AC2Activated carbon

Highly hydrophobic, low ash content, activated carbon used to remove or enrich very polar organic molecules from water.

JPMHLW official method for 1,4-dioxane analysis in water Pesticides,herbicides,esp.highlypolarsmallmolecules

Particle size: 85 m Quantity: 400 mg/cartridge

Sep-Pak Carbon Black/AminopropylCarbon black Aminopropyl silica

Two-layer sorbent bed used for pesticide cleanup in food matrices prior to GC analysis.

JPMHLW official methods for pesticides in food JPMHLW official method for propham

Particle size: 37105 m [carbon black, top layer] 55105 m [aminopropyl silica] Quantity: 500 mg of each sorbent, separated by frit Particle size: 37-105 m [carbon-black, top layer] 37-55 m [ PSA, bottom layer] Quantity: 500 mg of each sorbent, separated by frit

Sep-Pak Carbon Black/PSAPrimary-Secondary Amine Silica

Two-layer sorbent bed used for pesticide clean-up in food matrices prior to GC analysis. PSA provides alternative selectivity compared to aminopropyl.

JPMHLW official methods for pesticides in food.

* AOAC = Association of Official Analytical Chemists; ASTM = American Society for Testing and Materials [International]; CDFA = California Department of Agriculture; EPA = U.S. Environmental Protection Agency; JPMHLW = Japanese Ministry of Health, Labour and Welfare; JPMOE = Japanese Ministry of the Environment; NIOSH = National Institute for Occupational Safety and Health.

Matrix Key

Air

Petroleum

Biological

Soil

Water

Food

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Sample Preparation

15

SaMPLE PREPaRaTION Oasis Solid-Phase Extraction (SPE) ProductsA Breakthrough in SPE

IntroductionIn October 1977, Waters designed the first miniature cartridge columns (Sep-Pak cartridges) containing silica-based adsorbents for SPE. Demands for sample preparation led to the development of a speciallydesigned polymeric sorbent that performs optimally for reversed-phase SPE. T he Oasis HLB copolymer with unique Hydrophilic-Lipophilic Balance is unlike traditional SPE sorbents. Todays goals for modern solid-phase extraction (SPE) are faster throughput, higher recovery and reproducibility, and stronger retention and selectivity. Now SPE can outpace high-throughput techniques such as LC/MS/MS. Unique Water-Wettable Oasis HLB Copolymer T he Oasis HLB sorbent is a macroporous copolymer made from a balanced ratio of two monomers, the lipophilic divinylbenzene and the hydrophilic N-vinylpyrrolidone. It provides reversed-phase capability with a special polar hook for enhanced capture of polar analytes and excellent wettability.

High and Consistent RecoveriesOasis sorbents are water-wettable maintaining high retention and capacity for a wide spectrum of analytes, especially when the SPE column runs dry. W hen the sorbent pores dry out, the c hromatographic retention (capture) of the analytes is reduced, resulting in poor recovery. Traditional, silicabased C18 sorbents can easily dry out, especially on a vacuum manifold if a particular cartridge flows quickly and allows air to be drawn in. Oasis sorbents maintain proper wetting for more consistent performance (especially important for 96-well plates). Even if air passes through, the Oasis pores do not dry out.

Pore Dewetting Mechanism of Sorbent Pores (Silica Based C18)

N-VINYLPYRROLIDONE

DIVINYLBENZENE

Analytes properly retained in pore

Analytes not retained

Properly Wetted PoreHYDROPHILIC-LIPOPHILIC BALANCE Specific Surface Area: 810 m2/g, Average Pore Diameter: 80 Total Pore Volume: 1.3 cm3/g, Average Particle Diameter: 30 mm or 60 mmCurrent Oasis Patents:Patent No. 5,882,521 (1996), Patent No. 5,976,376 (1998), Patent No. 6,106,721 (1999), Patent No. 6,254,780 (2001) Patent No. 6,322,695 (2001), Patent No. 6,468,422 (2002), Patent No. 6,726,842 (2004), Patent No. 6,773,583 (2004) Patent No. 6,723,236 (2004), Additional Patents Pending

Dewetted PoreIf air passes through traditional C18 cartridge the pores dry out. When sample is loaded, the analytes are not retained resulting in poor recoveries.

16

Oasis Solid-Phase Extraction (SPE) Products

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SaMPLE PREPaRaTION

Effect of Drying on RecoveryOasis HLB Versus C18 SorbentsNo impact of sorbent drying on HLB High, consistent recoveryOasis HLB Cartridge (30 mg) 100 80 % Recovery 60 40 20 0 Procainamide Acetaminophen Ranitidine Propranolol Doxepin % Recovery 100 80 60 40 20 0 C18 Cartridge (100 mg)

Exceptional Batch-to-Batch ReproducibilityBecause of poor stability at pH extremes and relatively low ionic capacity traditional silica-based, mixed-mode sorbents dont have long-term batchto-batch reproducibility and therefore require reservations of specific lots of sorbent for large projects. Oasis sorbents have demonstrated excellent longterm, batch-to-batch reproducibility for over ten years. As a result of careful process design and stringent quality controls, a new standard has been set in batch-to-batch and lot-to-lot reproducibility for SPE sorbents. T he Oasis family of sorbents and devices are manufactured in a Waters ISO 9002 registered facility in compliance with cGMP guidelines of the U.S. Food and Drug Administration for class 1 medical devices.8 Min.

0

5 Drying Time

10 Min.

0

4 Drying Time

Multiple batches of each Oasis HLB, MCX, and MAX have been successfully used on validated bioanalytical assays in a regulated laboratory environment.

Oasis HLB 1 cc/30 mg and 1 cc/100 mg C18 cartridges were conditioned on a Waters vacuum manifold. When the methanol reached the top of the upper frit in each cartridge, vacuum was maintained for different times to vary the cartridge drying time. The SPE protocol was then continued. The data shown are the average of three replicate extractions.

Batch-to-Batch Reproducibility of Oasis HLB SorbentNo batch reservations needed: consistently high recoveries over more than 10 years of production105

T he variable recoveries seen with the C18 sorbents, due to the drying out effect, are often the cause for retests, reducing laboratory productivity. In some laboratories 10% of samples are reteststhis can be reduced using Oasis sorbents. Also, Oasis sorbents retain polar compounds far better than bonded silica SPE sorbents. Note the poor recovery of the polar analyte Acetaminophen for C18. Oasis sorbents work especially well when you need to capture metabolites (see figure above).

% Recovery

95 85

Recovery for:

Procanamide 1.90% RSD Ranitidine 1.68% RSD Acetaminophen 1.65% RSD

Oasis SPE ApplicationsOasis products come in a full range of device formats to meet your SPE requirementsElution plates, on-line columns, 96-well plates, and single-use cartridges. Try Oasis and successfully meet your SPE c hallenges. Download your free Oasis Applications Notebook at www.waters.com/oasis

High CapacityUse Less SorbentWhen transferring methods from a C18-bonded phase to Oasis products, keep in mind the greater capacity of the Oasis sorbent. The Oasis sorbent has 2-3X more surface area and shows a dramatic increase in k values compared to silica-based C18, This reduces breakthrough potential. In addition, you may be able to use 2/3 less sorbent than you would with C18 (30 mg Oasis HLB gives equivalent capacity to 100 mg C18). Higher Retention Means Greater Capacity, No BreakthroughRetention Factor (k) Comparison

H O O HO O H O N N CH CH N3

C18 CartridgeSalicylic acid TheobromineN

Oasis HLB Cartridge

Literature ReferencesOasis Sample Extraction Products Brochure, Literature Reference 720001692EN Oasis Elution Plate Brochure, Literature Reference 720000476EN Oasis WAX Sorbent for UPLC/MS Determination of PFOS and Related Compounds in Waters and Tissue, Literature Reference 720001871EN SPE Sample Preparation for UPLCMS Determination of Enrofloxacin (Baytril) in Chicken, Literature Reference WA43206 A Sensitive Method for the Determination of Endocrine-Disrupting Compounds in River Water by LC/MS/MS, Literature Reference 720001296EN

3X 2X 6X 15X0 50 100

3

H N H3C O HO OH HO

Acetaminophen Catechol

Topics in Solid-Phase Extraction. Part 1. Ion Suppression in LC/MS Analysis W hite Paper, Literature Reference 720001237EN Sample Prep Solutions Brochure, Literature Reference 720000848EN

Data shown were obtained with two 3.9 x 150 mm columns, each packed with one of the sorbents, operated under the same conditions: mobile phase: 20 mM potassium phosphate, pH 7.0/methanol (95/5 v/v); temperature: 30 C; flow rate: 1.0 mL/min; detection: UV @ 254 n

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17

SaMPLE PREPaRaTIONOasis MCX for Basic CompoundsOasis MCX Mixed-Mode Cation eXchange and Reversed-Phase Sorbent for High Selectivity and Sensitivity for Basic Compounds Obtain selective retention of basic drugs with cation-exc hange groups on the sorbent surface. T he Oasis MCX sorbent has a tightly controlled ion-exchange capacity (1 meq/g). T here are no silanol groups to complicate the retention mode or method development. T his novel, water-wettable, polymeric sorbent is stable from pH 0 to 14, making method development simple and fast. Drug/Sorbent Interactions on Oasis MCX SorbentHighly selective retention enables much stronger washes, resulting in very clean extracts.O MCX sorbent Mixed-mode Cation eXchange SO 3

Oasis MCX Retention Map for Basic Analytes Impact on Retention Factor (k) for a Base by Changing Mobile Phase pH in Cation-Exchange and Reversed-Phase Mode60 50 Retention (k) 40 30 20 10 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14

Base (Ionized)

Base (Unionized) Cation Exchange Reversed Phase

Mobile Phase pHNReversed-Phase Retention Cation-Exchange Retention Basic Analyte (Combined) Retention

SO 3

Reversed-phase interaction

Strong cationexchange mode Best retention at least 2 pH units below pKa of the analyte CH 3 O N H H OH Propranolol (basic drug)

+

CH 3

T his Retention Map plots the total k or capacity (retention) of a basic analyte relative to pH. Note that the total k, is the sum of the two retention mechanisms. At low pH, the analyte is charged and experiences maximum retention primarily from the ion-exchange mechanism, however, there is also a slight amount of reversed-phase contribution for the combined retention. If your goal is to capture basic analytes and then wash out interferences aggressively, the Load and Wash steps should be at low pH to obtain maximum capture. At high pH, the ion-exchange retention mechanism shuts-off because the analyte becomes un-ionized. Only reversed-phase retention is present, but since the analyte is now un-ionized, we get the maximum of the reversedphase retention. We can elute with a combination of high pH and high organic concentration. Generic Oasis MCX Method for Extraction of Basic CompoundsCondition: CH3OH Equilibrate: H2O Load: Spiked and acidified sample Wash 1: 2% HCOOH in H2O Elute 1(Wash 2): CH3OH Elute 2: 5% NH4OH in CH3OH Evaporate and Reconstitute: 20% CH3OH in H2O

Sulfonic-acid-cationexchange capacity: 1.0 meq/g

Since this ion-exc hange sorbent is synthesized from the reversed-phase Oasis HLB copolymer, it features two retention mec hanisms (cationexchange and reversed-phase) which can be manipulated very predictably (please refer to the Oasis MCX Retention Map).

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SaMPLE PREPaRaTIONOasis MAX for Acidic CompoundsOasis MAX Mixed-Mode Anion eXchange and Reversed-Phase Sorbent for High Selectivity and Sensitivity for Acidic Compounds T he Oasis MAX sorbent has a tightly controlled ion-exchange capacity of 0.3 meq/g ensuring reproducible SPE protocols for extraction of acidic compounds and metabolites from biological fluids. T here are no silanol groups to complicate the retention mode or method development. T his novel, water-wettable, polymeric sorbent is stable from pH 0 to 14, making method development simple and fast. Since this ion-exchange sorbent is synthesized from the reversed-phase Oasis HLB copolymer, it features two retention mec hanisms (anionexchange and reversed-phase) which can be manipulated very predictably (please refer to the Oasis MAX Retention Map, below).Reversed-phase retention Quaternary-amineanion-exchange capacity: 0.25 meq/g O CH 3

Drug/Sorbent Interactions on Oasis MAX SorbentHighly selective retention enables much stronger washes, resulting in very clean extracts.

O MAX sorbent Mixed-mode Anion eXchange

N

CH 2 N R+

CH 3 + CH 3

C4H9

S

COO

Strong anionexchange mode Best retention at least 2 pH units above pKa of the analyte

Suprofen (acidic drug)

Oasis MAX Retention Map for Acidic Analytes Impact on Retention Factor (k) for an Acid by Changing Mobile Phase pH in Anion-Exchange and Reversed-PhaseAcid (Ionized)

At low pH, the ion-exchange retention mechanism shuts-off because the analyte becomes un-ionized. Only reversed-phase retention is present, but since the analyte is now un-ionized, we get the maximum of the reversedphase retention. We can elute with a combination of low pH and high organic concentration. Generic Oasis MAX Method for Extraction of Acidic Compounds

60 50Retention (k)

40 30 20 10 0 1

Acid (Unionized) Anion Exchange Reversed Phase

Condition: CH3OH Equilibrate: H2O Load: Spiked acidified sample

2

3

4

5

6

7

8

9 10 11 12 13 14

Mobile Phase pHReversed-Phase Retention Anion-Exchange Retention Acidic Analyte (Combined) Retention

Wash 1: 5% NH4OH in H2O Elute 1 (Wash 2): CH3OH Elute 2: 2% HCOOH in CH3OH Evaporate and Reconstitute: 20% CH3OH in H2O

T he Retention Map plots the total k or capacity (retention) of an acidic analyte relative to pH. Note that the total k, is the sum of the two retention mechanisms. At high pH, the analyte is charged and experiences maximum retention primarily from the ion-exchange mechanism, however, there is also a slight amount of reversed-phase contribution for the combined retention. If your goal is to capture acidic analytes and then wash out interferences aggressively, the Load and Wash steps should be at high pH to obtain maximum capture.

As shown (next page), acidic compounds can be separated from basic and neutral compounds on the same Oasis MAX cartridge following the recommended protocol.

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19

SaMPLE PREPaRaTION

High Recovery of Acidic, Basic, and Neutral Compounds Using the Generic Oasis MAX Method Elute 1 (basic and neutral drugs)1

Elute 2 (acidic drugs)

2 I.S. 3 4 I.S.

AV

Sample Blank with I.S.0Column:

AV

Sample Blank with I.S.0 2 4 6 8 10 Min.

2

4

6

8Peak Identification 1. Nortriptyline 2. Secobarbital 3. Ketoprofen 4. Naproxen I.S. Butylparaben

10 Min.

Mobile Phase: Detector: Temperature: Flow Rate: Injection Volume:

SymmetryShield 3.5 m, 4.6 x 100 mm SymmetryShield Guard column, 5 m, 3.9 x 20 mm 20 mM potassium phosphate, pH 2.7/methanol, 52:48 v/v UV @ 214 nm 30 C 2 mL/min 10 L of extract with internal standard

Results (n=6) Drugs Nortriptyline Secobarbital Ketoprofen Naproxen g/mL 5 10 5 2.5 Type of Drug Basic (Elute 1) Neutral (Elute 1) Acidic (Elute 2) Acidic (Elute 2) % Recovery 94.34 88.53 91.64 103.43 % RSD 0.78 1.03 2.35 3.04

* 1 mL urine hydrolyzed (1M KOH 60 C, 15 minutes) and adjusted to pH 2 with phosphoric acid; diluted 1:1 with 10 mM sodium acetate, pH 2.

Literature ReferencesOasis Sample Extraction Products Brochure, Literature Reference 720001692EN Oasis Elution Plate Brochure, Literature Reference 720000476EN Topics in Solid-Phase Extraction. Part 1. Ion Suppression in LC/MS Analysis W hite Paper, Literature Reference 720001237EN Sample Prep Solutions Brochure, Literature Reference 720000848EN Oasis WAX Sorbent for UPLC/MS Determination of PFOS and Related Compounds in Waters and Tissue, Literature Reference 720001871EN SPE Sample Preparation for UPLCMS Determination of Enrofloxacin (Baytril) in Chicken, Literature Reference WA43206 A Sensitive Method for the Determination of Endocrine-Disrupting Compounds in River Water by LC/MS/MS, Literature Reference 720001296EN

PST T herapeutic Peptide Method Development KitT he PST T herapeutic Peptide Method Development Kit has been developed to simplify the process of sample preparation and LC method development for the analysis of therapeutic peptides in plasma. T he kit contains an Oasis PST Elution Method Development Plate, a PST 300 C18 reversed-phase column and the detailed screening protocol.

See Page 209

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SaMPLE PREPaRaTIONOasis WCX for Strong Basic CompoundsHigh Selectivity and Sensitivity for Strong Basic CompoundsHighly selective retention enables much stronger washes, resulting in very clean extracts.

Generic Oasis WCX Method for Strong Basic CompoundsCondition: CH3OH Equilibrate: H2O Load: Spiked acidified sample

O WCX sorbent Weak Cation eXchange

N

COO Weak cation-exchange interaction CH3 H 3C Reversed-phase interaction O O CH3 N + CH3

Wash 1: 5% NH4OH in H2O Elute 1 (Wash 2): CH3OH Elute 2: 2% HCOOH in CH3OH Evaporate and Reconstitute: 20% CH3OH in H2O

CH3

Carboxylic-acid-cationexchange capacity: 0.75 meq/g

T he Oasis WCX (Weak Cation eXchange) SPE material was developed to provide better sample preparation for strong bases and quaternary amines. T he retention mechanism is mixed mode, both ion exchange and reversed phase, which improves retention for all types of basic analytes, especially strong bases. Recovery Data for T hree Basic Compounds from Oasis WCX vs. Silica-Based WCX Products Excellent Recovery for Quaternary Amines as well as Polar and Hydrophobic BasesColumn: Mobile Phase A: Mobile Phase B: Gradient: Flow Rate: Injection Volume: XTerra MS C18 IS 2.1 x 20 mm, 3.5 m 10 mM NH4HCO3, pH 10 MeOH with 10 mM NH4HCO3, pH 10 5 to 95% B in 3 min 0.4 mL/min 10 L Instruments: Waters 2777 Sample Manager, and Waters 1525 binary HPLC pump, Waters Quattro Ultima (ESI in MRM)

(a)Recovery (%)

Oasis WCX 96-well Elution Plate120.0 100.0 80.0 60.0 40.0 20.0 0.0 0.25 Loading volume (mL) 1.00

Protriptyline (H) Valethamate (Q) Atenolol (P)

MRM 266.9 > 144.9

Atenolol 10 ng/mL 101% Recovery

(b)Recovery (%)

120.0 100.0 80.0 60.0 40.0 20.0 0.0

Silica-Based WCX (CBA) 10 mg, 96-well Plate

MRM 306.1 > 218.9

Valethamate 10 ng/mL 106% Recovery

Protriptyline (H) Valethamate (Q) Atenolol (P)

MRM 264 > 191.1

Protriptyline 10 ng/mL 102% Recovery

0.25 Loading volume (mL)

1.00

SPE recoveries for three types of bases versus volume of spiked saline solution loaded onto (a) the Oasis WCX sorbent and (b) a commercially available silica-based WCX sorbent. H is hydrophobic base, Q is quaternary amine, and P is polar base.

LC/MS/MS chromatograms of the analytes (polar base, quaternary amine, and hydrophobic base) extracted from rat plasma with Oasis WCX 96-well Elution plate. The SPE recoveries were calculated by comparison with postextracted spiked plasma samples.

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21

SaMPLE PREPaRaTIONOasis WAX for Strong Acidic CompoundsHigh Selectivity and Sensitivity for Strong Acidic CompoundsHighly selective retention enables much stronger washes, resulting in very clean extracts.

Generic Oasis WAX Method for Strong Acidic CompoundsCondition: CH3OH Equilibrate: H2O

O WAX sorbent Weak Anion eXchange

N

+ NH

Load: Spiked and acidified sample+ NH 2 Weak anion-exchange interaction

Wash 1: 2% HCOOH in H2O Elute 1(Wash 2): CH3OH Elute 2: 5% NH4OH in CH3OH Evaporate and Reconstitute: 20% CH3OH in H2O

SO 3 Reversed-phase interaction Piperazine-anion-exchange capacity: 0.6 meq/g

T he Oasis WAX (Weak Anion-eXc hange) SPE material was developed to provide sample preparation for strong acidic compounds. T he retention mechanism is mixed mode, both ion exchange and reversed phase, which improves retention for strong acidic compounds. Recovery Data for T hree Acidic Compounds from Oasis WAX 1 cc, 30 mg Cartridge100 90 80 % Recovery 70 60 50 40 Saline. pH = 7 Saline, pH = 1.3 pH 1.3 Rat Plasma

Excellent Recovery for Strong Acid: Camphorsulfonic AcidHPLC Conditions: Column: Mobile Phase A: Mobile Phase B: Flow Rate: Gradient: MS/MS Conditions: Waters Quattro Premier ESISource Temp: Desolvation Temp: Cone Gas Flow: Desolvation Gas Flow: Collision Cell:CH3

2-Naphthalenesulfonic acid Ketoprofen Salicylic Acid

SPE recoveries for three types of acidic compounds spiked in saline pH 7 and 1.3, and in rat plasma pH 1.3. Recovery Data of T hree Acidic Compounds for Oasis WAX vs. Silica-Based WAX Products

Injection Volume: Temperature: Instrument:

SunFire C18 2.1 x 20 mm IS, 3.5 m H2O with 10 mM CH3COO-NH4+, pH 7 MeOH with 10 mM CH3COO-NH4+, pH 7 0.4 mL/min Time Profile (min) %A %B 0.0 95 5 3.0 5 95 4.0 5 95 4.1 95 5 5.0 95 5 10 L Ambient Waters 2777 Sample Manager, Waters 1525 Binary HPLC Pump and Waters Quattro Premier

150 C 350 C 50 L/Hr 600 L/Hr 2.2e-3 bar (Ar gas)CH3

O CH2SO3H

(1S)-Camphor-10-sulfonic acid MW 232.3, C10H16O4S MRM Transition m/z 231.1 _ 79.8 Cone (V) 60 CID (eV) 30

100100 80 % Recovery 60 40 20 0 DEA PSA Oasis WAX 1 cc/ 1 cc/ 1 cc/ 100 mg 100 mg 30 mg

1.59

Breakthrough at Load Step

Recovery at Elution Step

Concentration: 10 pg/L 108.8% recovery

2-Naphthalenesulfonic acid Ketoprofen Salicylic AcidOasis WAX 1 cc/ 30 mg DEA PSA 1 cc/ 1 cc/ 100 mg 100 mg

% 1.29 1 100 1.59

2.75 3.10

4.20

4.33

Spiked Saline, pH = 7

Concentration: 50 pg/L 114.4% recovery

%

SPE recoveries for three types of acidic compounds versus volume of spiked saline solution and rat plasma loaded onto (a) the Oasis WAX sorbent and (b) a commercially available silica-based WAX sorbent.

0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5 min

22


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SaMPLE PREPaRaTIONOasis HLB for Reversed-Phase SPEThis generic, 1-D method (1-Dimensional only the organic strength is changed) has proven useful for a wide variety of compound types and may be the only protocol required, reducing method development time.O N

Recommended Generic Oasis HLB SPE Method (1-D)*SO 3

O

N

SO 3

CH + CH 2 N

Condition: 1 mL CH3OH Equilibrate: 1 mL H2O

R+

CH

Oasis HLB Sorbent Oasis HLB sorbent makes it fast and easy to develop sample preparation methods that deliver high, reproducible recoveries especially suited to LC/MS/MS analysis by providing the required selectivity and sensitivity. A Simple and Fast Generic SPE Reversed-Phase Protocol for Rapid Method Development for a Wide Range of Compounds (1-D) Limitations of traditional silica-based sorbents make you evaluate several different bonded phases and brands to obtain acceptable results. With Oasis HLB, acidic, basic, and neutral compounds, whether polar or nonpolar, can be isolated reproducibly (RSDs 85%), using the same simple SPE protocol (see below).

Load: 1 mL spiked sample** Wash (Mild Wash): 1 mL 5% CH3OH in H2O Elute (Strong Elution): 1 mL CH3OH Evaporate and Reconstitute: 40 C/under nitrogen stream 200 L mobile phase* Volumes are given for the Oasis HLB cartridge 1 cc/30 mg

Sample Pretreatment SuggestionApplying one or more of the following steps before loading your sample may improve your results: Dilute sample 1:1 with buffer to improve flow during loading ** Add 20 L (H3PO4) to disrupt drug-protein interaction/ binding Dilute 1:1 or greater with 0.1 N HCl or other acids Filter through 0.45 m membrane Centrifuge @ 3000 rpm

One Simple Procedure: Many Applications on a Universal SorbentReduced Methods Development Time

sed sorbents With silica-ba need several (C18) you would to t chemistries differen ults ac hieve the res shown here

100

Acids Neutrals Bases n=6 RSDs < 3.5%

80

60 % Recovery

40

1-D MethodSeventeen drugs with a wide range of polarities were spiked in porcine serum, then extracted with 1 cc/30 mg Oasis HLB cartridges, using the same SPE method. (20 L/mL H2PO4 was used to disrupt drug-protein binding)

20

0pr of Na en pr ox Sa en lic yl ic Ac Su id lfa Su diaz lfa i m ne er Ac az in et e am in op he Th n eo br Pa omi ra ne za nt hin Th e eo ph yl lin Ca e ffe Pr in oc e ain am id Ra e ni tid Ox ine yc od on Pr e op ra no Na lol ltr ex on Sa e lb ut am ol Do xe pin Ibu

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23

SaMPLE PREPaRaTION

Oasis 2x4 Methodology T he Oasis 2x4 Method is a simple, logical approach to the selection of an SPE sorbent and protocol. Two protocols and four sorbents provide the flexibility to extract acids, bases, and neutrals with high SPE recoveries while removing matrix components that may interfere with analysis. Follow the simple steps outlined in this flow c hart to ac hieve high recoveries and the cleanest extracts:For Bases pKa 2-10: Use Oasis MCX

Oasis 2x4 MethodOnly 2 protocols and 4 sorbents to analyze all types of compounds: acids, bases, and neutrals

Characterize your analyte [Neutral, Acid, or Base, pKa] Select one of the four Oasis sorbents Apply the indicated Protocol [1 or 2] Determine SPE recoveries by LC analysis

For Strong Acids pKa 10: Use Oasis WCX

Apply Protocol 1 Prepare Sample Condition/Equilibrate Load Sample Wash: 2% HCOOHWeaker Acids

Apply Protocol 2 Prepare Sample Condition/Equilibrate Load Sample Wash: 5% NH4OHNeutrals

Elute 1: 100% CH3OH Elute 2: 5% NH4OH in CH3OH

Elute 1: 100% CH3OH Elute 2: 2% HCOOH in CH3OH

Weaker Bases

Base

Strong Acid

Acid

Strong Base

Note that neutral analytes can be isolated from any of the four sorbents in the Elute 1 step of either protocol. Choose the particular ion-exchange sorbent that is best at removing specific matrix interferences. A good example of this is shown below. Choosing Optimum Sorbent and Protocol for Neutral Compounds Example: Prednisone in PlasmaSPE Recovery100

Oasis sorbent selection plate and cartridge kits enable rapid development of SPE methods for LC/MS analysis. Having all four Oasis ion-exc hange sorbents [MCX, MAX, WAX, and WCX] in a single plate is convenient for scouting the best ways to accomplish efficient isolation of unknowns, zwitterionic compounds, or mixtures of analytes with different retention/ elution properties.

Cleanliness of 100% MeOH Elute 1

Monitoring 5 Phospholipid MRM Transitions

0

50

100%100 %

Protein Precipitation

%

2.52 2.60

3.26 3.16 3.08

1: MRM of 5 Channels ES+ TIC 2.55e7

100% Phospholipids 2% Phospholipids

Oasis MCX Protocol 12.50


100 %

Oasis MAX Protocol 22.522.60


9% Phospholipids 11% Phospholipids

100 %

Oasis WAX Protocol 12.50 2.60


Oasis Sorbent Selection Plate: Evaluating Oasis 2x4 Method for Cephalexin0 SPE Recovery 50 100%

100 %

Oasis WCX Protocol 22.52 2.60 1.75 2.00 2.25 2.50 2.75 3.00 3.25

1: MRM of 5 Channels ES+ TIC 2.55e7 3.50 3.75 4.00 Min

12% PhospholipidsO O H H O H OH OH

O NHpKa = 7.3

H H N H O N HO

0

Oasis MCXProtocol 1 Elute 22

S

Recovery is high on all four Oasis sorbents, but removal of phospholipids, a primary cause of matrix ion suppression, is best on Oasis MCX. Protein precipitation is a poor choice for sample preparation prior to LC/MS/MS analysis.

Oasis WAXProtocol 1 Elute 1

Prednisone

Oasis MAXProtocol 2 Elute 2

pKa = 2.6

O

Oasis WCXProtocol 2 Elute 1

Cephalexin [Cephalosporin antibiotic] A difficult amphoteric analyte

Aliquots of prepared sample processed using Oasis 2x4 Method protocol designated for each of 4 sorbents. Eluates from Elute 1 and Elute 2 steps analyzed by LC/MS/MS. Clearly, Oasis MCX is the sorbent of choice.

24


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SaMPLE PREPaRaTION

Oasis 2x4 Method Proof-of-Concept To demonstrate the logic, simplicity, and effectiveness of the Oasis 2x4 Method, five rat plasma samples were prepared, each containing one of these characterized test analytes:

and the Oasis ion-mixed-mode sorbent designated by the Oasis 2x4 Method for the corresponding sample type. LC/MS/MS analysis was used to determine SPE recoveries. T he neutral analyte was processed on all four sorbents, as shown on the previous page. Of the four method options, Oasis MCX with Protocol 1 proved superior at removing nearly all the phospholipids, eliminating this major source of matrix effects, a known cause of ion suppression, loss of sensitivity, and inaccurate quantitation in LC/MS analysis. Essentially, quantitative recovery and excellent cleanup efficiency were ac hieved for eac h of t he ionic or ionizable test analytes w hen t he recommended Oasis 2x4 Method sorbent/protocol combination was used. T hese results are shown in the four figures below.

Imipramine, a base pKa of conjugate acid = 9.4 Ibuprofen, an acid pKa = 5.2 Decanesulfonic acid, a strong acid pKa < 0.5 Valethamate, a quaternary amine [strong base] pKa of conjugate acid > 12 Prednisone, a neutral compound

Each plasma sample was diluted [1:1, v/v] and acidified with phosphoric acid [4% in water]. Respective aliquots were then processed using the protocol Oasis 2x4 Method Test on MCX: Base IsolationOasis MCXFor Bases pKa 2 1 0 : Use Oasis MCX

Oasis 2x4 Method Test on WAX: Strong Acid IsolationOasis WAXFor Strong Acids pKa < 1: Use Oasis WAX

Protocol 1

Protocol 1

pKa 85.95 4.16e6

SPE Recovery: 94% n=8, 3-6% RSD

pKa ~6O S ODecanesulfonic acid [pKa 79.8 2.85e3

N N

O Na

+

Imipramine [pKa = 9.4]

0

1

3

5 Min.

0

1

3

5 Min.

Oasis 2x4 Method Test on MAX: Acid IsolationOasis MAXFor Acids pKa 28: Use Oasis MAX

Oasis 2x4 Method Test on WCX: Strong Base IsolationOasis WCXFor Strong Bases pKa > 10: Use Oasis WCX

Protocol 2

Protocol 2

pKa ~18

Ibuprofen

Valethamate

SPE Recovery: 100% n=6, 6-13% RSD2.63

pKa ~5O Br

SPE Recovery: 106% n=4, 9-13% RSD2.71

OH OIbuprofen [pKa = 5.2]0 1 3

MRM ES+ 205 > 161 1.34e4

O

N

+

MRM ES+ 306.2 > 163 2.10e6

5 Min.

Valethamate [pKa >12]

0

1

3

5 Min.

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25

SaMPLE PREPaRaTIONOasis Elution PlatesWaters Innovative SPE Technology

Elute in as little as 25 L No evaporation and reconstitution Ideal for small sample volumes Up to a 15x increase in concentration

Now you can confidently perform SPE cleanup and analyte enrichment of very small sample volumes (10-25 L) up to a maximum of 375 L. T he Waters Oasis Elution plate combines patented plate design, proven Oasis chemistries, and straightforward protocols that deliver high analyte recovery and clean extracts in elution volumes as low as 25 L.

Reproducible elution in only 25 L enhances and streamlines SPE methods. SPE can be performed on very small sample volumes. Sample enrichment increases as much as 15-fold. A time-consuming step for eluate concentration by evaporation is no longer neededneither is evaporation/reconstitution if the eluting solvent is properly chosen to be compatible with the LC/ MS mobile phase. T he innovative features of Oasis Elution plates enable sensitive, robust, reproducible results without evaporation and reconstitution. Analytical performance, in terms of sensitivity, selectivity, and ruggedness, is superior. Comparison of Elution vs. Disk TechnologyElution Technology Disk Technology

Narrow and Tall Sorbent Bed Ratio Bed H/D = 1.15

Wide and Short Sorbent Bed

Ratio Bed H/D = 0.13

Oasis Elution Technology vs. Disk Technology T here are two predominant low-elution SPE technologies in the marketplace today: SPE Disk and Oasis Elution. T hese technologies differ greatly in performance primarily because of three features: aspect ratio, holdup volume and elution volume. T here is a correlation between a sorbent beds aspect ratio [ height to diameter, H/D] and its performance in an SPE device. An H/D ratio less than 1.0 often compromises extraction efficiency. In an SPE disk design (e.g. membrane/glass-fiber disk), a small mass of sorbent is embedded in a thin support structure having good flow properties but a small aspect ratio, H/D=0.13. In contrast, the Oasis Elution tec hnology uses an internally tapered well, packed with high capacity Oasis sorbent, having an aspect ratio, H/D=1.15, nearly 9x that of SPE disk tec hnology. An Oasis Elution plate well functions more like a c hromatography column, minimizing loss of analyte during critical steps in the SPE procedure and enhancing overall extraction efficiency. A small holdup volume is necessary to minimize elution volume. SPE disks have holdup volumes in the 3565 L range, compared to that for an Oasis Elution bed plus frits which measures as little as 15 L. Having 24x less holdup volume minimizes elution volume and reduces analyte loss during elution, improving both recovery and precision. Minimizing SPE elution volume is critical for SPE performance, recovery, and precision. Normal elution volume range for an Oasis Elution plate is only 2550 L, far lower t han t he 75300 L typical for SPE disk tec hnology.

Vacuum

Vacuum

Recovery Comparison of Elution vs. Disk TechnologyT he innovative features of the Oasis Elution plates enable sensitive, robust, reproducible results without evaporation and reconstitution.Oasis Elution Technology >85% recovery in 25 L25 L

Disk Technology Minimum of 100 L needed for best result

75 L

75 LElution Volume

75 L

100 80 % Recovery 60 40 20 0 Oasis HLBElution plate

Brand E C18

Brand E UR

Brand S C18

100 L Spiked Saline acetaminophen practolol N-acetylprocainamide betamethasone caffeine naproxen amitriptyline propranolol

26


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SaMPLE PREPaRaTION

Oasis 2x4 Method Optimized for Elution Plate T he proven Oasis 2x4 method elution solvent is optimized to accommodate the elutropic requirement of the small elution volume. Methanol is good as a generic elution solvent, but is often not strong enough for 25 L elution volumes. T he elution solvent recommended to be used with the Elution plate must possess a high enough elutropic strength to fully elute analytes in small volumes, and be appropriate for a diverse set of analytes. T he recommended elution solvent for the Oasis 2x4 method optimized for the Elution plate format is 60% CH3CN:40% CH3OH with a modifier. T his was chosen as a starting point as it meets all of the above criteria. Follow the simple steps outlined in this flow chart to achieve high recoveries and the cleanest extracts:

For Bases pKa 2-10: Use Oasis MCX

For Strong Acids pKa 10: Use Oasis WCX

Apply Protocol 1 Prepare Sample Condition/Equilibrate Load Sample Wash: 2% HCOOH Elute 1: 100% CH3OH Elute 2: 5% NH4OH in 60:40 CH3CN:CH3OHNeutrals

Apply Protocol 2 Prepare Sample Condition/Equilibrate Load Sample Wash: 5% NH4OH Elute 1: 100% CH3OH Elute 2: 2% HCOOH in 60:40 CH3CN:CH3OH

Characterize your analyte [Neutral, Acid or Base, pKa]. Select one of the four Oasis sorbents. Apply the indicated Protocol [1 or 2]. Determine SPE recoveries by LC analysis.

Base

Strong Acid

Acid

Strong Base

Oasis 2x4 Method Proof-of-Concept Recovery Study: To demonstrate the logic, simplicity, and effectiveness of the Oasis 2x4 method, five samples of rat plasma were prepared, each spiked with one of the previously characterized test analytes shown below: Imipramine: pKa = 9.4 (Base) Ibuprofen: pKa = 5.2 (Acid) Valethamate: pKa >12 (Quaternary Amine) Nonafluoropentanoic Acid: pKa 12, 2 ng/mL

O

N+

Br O H H O

O

OH OH

H

Prednisone (N) 10 g/mL

O F HO

F

F F F OIbuprofen (A) pKa = 5.2, 200 ng/mL

OH

F F F F

Nonafluoropentanoic Acid (SA) pKa 95% collection efficiencies for all aldehydes at flows of up to a liter per minute. And, you only need to use one cartridge no breakthrough bed is necessary. Low Pressure DropUse with Portable Personal Sampling Pumps Large particle size and higher porosity frits make the X PoSure cartridge compatible with personal sampling pumps. Easy-to-Use Sample, elute and shoot, its that easy. Youll never have to break open and manipulate a glass tube again. And because the cartridges are made from high density polyethylene (HDPE), breakage is not a concern. T he figure on t he right s hows two traces. An actual cartridge blank demonstrating extremely low background levels and as an actual laboratory air sample. X PoSure Aldehyde Sampler CartridgeDescription XPoSure Aldehyde Sampler Cartridge Qty. 20/box Part No. WAT0472052

Low-Level Example: Aldehyde Profile from Laboratory Air1 4

1. 2. 3. 4. 5. 6.

DNPH Formaldehyde-DNPH Acetalhyde-DNPH Acetone-DNPH Butanone-DNPH Isoval

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