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Waters Quality Parts, Chromatography Columns and Supplies Catalog

2011/2012

Waters Chrom

atography Columns and Supplies Catalog

2011 / 20122011 Waters Corporation. Printed in the U.S.A.720003820EN

SaleS OfficeS

Australia Waters Australia Pty. LimitedUnit 3, 38-46 South StreetP.O Box 84Rydalmere BC NSW 2116Tel: 2 9933 1777Fax: 2 9898 1455

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CanadaWaters Limited6427 Northam DriveMississaugaOntario L4V 1J2Tel: 800 252 4752Fax: 905 678 9237

China Waters Technologies (Shanghai) LimitedBlock 41, Room 01 1387 Zhangdong RoadPudong New DistrictShanghai 201203, PR ChinaTel: 86 21 6879 5888Fax: 86 21 6879 4588

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India Waters India Pvt. Ltd.36A, II PhasePeenya Industrial ParkBangalore 560058Tel: 91 80 2 837 1900Fax: 91 80 2 839 2157

IrelandWaters Chromatography Ireland Ltd., Unit 3.1 Woodford Business Park Santry, Dublin 9, IrelandTel: 353 1 448 1500Fax: 353 1 448 1510

Italy Waters S.P.A.Via Achille Grandi, 2720090 Vimodrone MITel: 39 02 265 0983Fax: 39 02 250 1827

JapanNihon Waters K.K.Dai-5. Koike Bldg.1-3-12 Kita-shinagawaShinagawa-ku, Tokyo 140-0001Tel: (81) 3 3471 7191Fax: (81) 3 3471 7118

Korea 4th Floor KICOX Venture Center 188-5 Guro-Dong, Guro-Gu Seoul 152-050 Korea Tel: (82) 2 6300 4800 Fax: (82) 2 6300 4830

Mexico Waters S.A. DE C.V.Moras No 822 Del Benito Juarez , Col Acacias C.P. 03230, Mexico D.F. Tel: 5255 5524 7636Fax: 5255 5524 9376

The Netherlands Waters Chromatography B.V.Florijnstraat 19Postbus 3794870 AJ Etten-LeurTel: 31 76 508 7200Fax: 31 76 508 7280

Norway Tel: 47 63 84 60 50Fax: 47 63 84 00 10

Poland Waters Sp z o.o ul. Lektykarska 25 m 21 01-687 Warszawa Tel: (48) 22 6393000 Fax: (48) 22 6393009

Puerto Rico Waters Corp PR BranchAve. Gautier Benitez #230Edif. Video Avenue 2nd FloorCaguas,PR 00725Tel: 787 747 8445Fax: 787 747 8448

Singapore Waters Asia Limited1 Science Park Road#02-01/06 The CapricornSingapore Science Park IISingapore 117528Tel: 65 6593 7100Fax: 65 6873 0733

SpainWaters Cromatografa, S.A. Ronda Can Fatj, 7-A Parc Tecnolgic del Valls 08290 Cerdanyola del Valls Barcelona Direct: 902 254 254Tel: 34 936 009 300Fax: 34 936 009 360

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SwitzerlandWaters AG Tfernstrasse 4Postfach5405 Baden-DttwilTel: 41 56 676 70 00Fax: 41 56 676 70 49

Taiwan Waters Asia Ltd., Taiwan Branch11F-2, No.90, Sec. 1, Jianguo N. Rd., Zhongshan Dist., Taipei City 10491, Taiwan, R.O.C.Tel: 886 2 2501 9928 Fax: 886 2 2501 9228

United Kingdom Waters U.K. Ltd.730-740 Centennial CourtCentennial Park, Elstree,Hertfordshire WD6 3SZTel: 44 208 238 6100Fax: 44 208 207 7070

All Other Countries

Waters Corporation34 Maple StreetMilford, MA 01757 USATel: 508 478 2000 800 252 4752Fax: 508 872 1990

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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

benchmark 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 world-

wide offices and interact with our highly trained account specialists.

Michael J. Yelle

Senior Director, Chemistry Commercial Operations

Waters Corporation Milford, MA, U.S.A.

http://www.waters.com/catalog

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Sample Preparation

How to Order

Sample Vials and Accessories

Filtration

ACQUITY UPLC Columns and Consumables

Analytical Chromatography

Preparative Chromatography

Supercritical Fluid Chromatography (SFC)

Bioseparations and Analyses

Size-Exclusion Chromatography GPC & GFC

Alphabetical Index

Supplies and Hardware

Part Number Index

CONTENTS

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.

Cover Image:

WhaTS NEW

Page 51

The 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.

Waters Positive Pressure-96 Processor

Page 53

Ostro 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 techniques, the Ostro

96-well plate allows for more sensitive analyses, increased sample

throughput, and reduced instrument downtime.

Ostro Sample Preparation Products

Page 91

CSH Technology is the next evolution of UPLC and HPLC particle

technology. It affords outstanding batch-to-batch reproducibility,

extended pH stability and seamless transferability between UPLC,

analytical HPLC and Preparative HPLC system platforms.

The ACQUITY UPLC CSH column family provides separation scientists

with a wider range of selectivities as well as industry-leading

performance in simple, LC/MS-friendly mobile phases.

ACQUITY UPLC CSH Columns

XSelect HPLC columns are the next evolutionary step in Waters

LC column technology. Based on Charged Surface Hybrid (CSH)

Technology, 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).

XSelect HPLC Columns

Page 118

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Whats NeW

Page 196

Combining state-of-the-art media manufacturing with the industry-

leading column technology, Viridis Supercritical Fluid Chromatography

(SFC) columns bring a new level of reproducibility to the world of

laboratory-scale purification.

Viridis Supercritical Fluid Chromatography (SFC) Columns

Page 7

Waters Online Order Center streamlines the order process by placing

product information, availability, technical 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.

Online Order Center and Digital Catalog

Page 217

ACQUITY UPLC analytical size-exclusion chromatography (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 such as

therapeutic mAbs.

ACQUITY UPLC BEH200 SEC Columns

Page 223

The 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.

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

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To view and download our latest literature, simply visit

www.waters.com

and enter the literature code in the search field (top right)

Analytical Columns Wall ChartLit. Code 720002241EN

Prep OBD Wall ChartLit. Code 720002117EN

Sorbent Selection Guide for SPE Wall Chart

Lit. Code 720002007EN

NEW ACQUITY UPLC CSH and XSelect HPLC Columns Brochure

Lit. Co de 720003495EN

ACQUITY UPLC Columns BrochureLit. Code 720001140EN

ACQUITY UPLC Column Strategies BrochureLit. Code 720003271EN

XBridge HPLC Columns BrochureLit. Code 720001255EN

SunFire HPLC Columns BrochureLit. Code 720000875EN

Prep OBD Columns BrochureLit. Code 720002336EN

Atlantis Columns BrochureLit. Code 720002861EN

NEW Viridis SFC Columns BrochureLit. Code 720003326EN

Oasis Sample Extraction Products Brochure


Sample Vials and Accessories Brochure


LITERaTURE ShOWCaSE

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Waters Vials Selectorwww.waters.com/vialsselector

Waters Carbohydrate Method Selection Tool

www.waters.com/amidemethod

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

NEW Waters Plate Selectorwww.waters.com/plateselector

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

NEW Waters Filter Selectorwww.waters.com/filterselector

Environmental Analysis Catalog


Food Testing Solutions Brochure


Environmental Analysis Solutions Brochure


Oasis Sample Preparation Application Notebook


Environmental Chromatography Methods Guide


NEW Ostro Sample Preparation Products,


Waters DisQuE Products for QuEChERS,


PoraPak RXN for Post Synthesis Cleanup Brochure


Sample Preparation Solutions Brochure


Bioseparations and Analyses Catalog


LITERATURE SHOWCASE

http://www.waters.com/vialsselectorhttp://www.waters.com/amidemethodhttp://www.waters.com/selectivitycharthttp://www.waters.com/plateselectorhttp://www.waters.com/prepcalculatorhttp://www.waters.com/filterselectorhttp://www.waters.com

Beginners Guide to UPLC 715002099

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 978-1-879732-05-6

Quest for Ultra Performance in Liquid Chromatography 715002098

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 UPLC Tec hnology principles. Scientists will gain the confidence to apply this knowledge in ways that enhance analytical productivity, streamline workflow, and advance scientific progress within their organizations.


Beginners Guide to Liquid Chromatography 715001531

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. This primer is suitable for younger science students as well as professionals new to LC.


NEW Comprehensive Guide to HILIC 715002531

NEW Comprehensive Guide to HILICHydrophilic Interaction ChromatographyT his technology primer is designed to provide the reader with the basic insight of how to be successful with hydrophilic interaction chromatography by understanding how the technique works, the parameters that impact retention and selectivity, as well as the practical considerations necessary to successfully implement HILIC within a c hromatographic strategy.


The Mass Spectrometry Primer 715001940

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.


Waters 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.

www.waters.com/primers

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LITERATURE SHOWCASE

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How to order

Waters Online Order Center

An Improved Online Shoping Experience At Waters.comWaters.com is a convenient interface for both purchasing/procurement departments and scientific users to research, compare and purchase 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.

[ Benefits of e-Commerce ]

Reduction of paper transactions

Stock availability in real time

Multiple Payment options

Streamline internal process

e-Order /e-Invoice

Lower transaction costs

Shorter order cycle time

High Compliance levels

Ease of finding products

Account specific pricing

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Quick Order

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

How to Buy Service Plans, Instruments, or Software

Options available for pricing and quotations.

Ordering FAQs

Frequently 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 Cart

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

Freight Policy

Delivery time options and freight charges in each country.

Offers & Discounts

Look through all offers and discount codes available before you buy.

Technical Support (iRequest)

Online support for technical queries on instrument/software.

Graphical Parts Locator

Exploded instrument diagrams help you locate specific spare parts quickly.

Order History

Find previous ordering information.

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

[ The Order Center Process ] Step 3 Buy online

Add discount coupons if available

Automatically applies your company discount at checkout

Choose shipping method

Choose credit card payment (if available)

Step 2 Register to buy online

Step 1 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

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W hether researching product information or making online purchases, 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 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.

How do I find chemistry related consumable products when I dont know what I am looking for?

There 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

b. 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 >

a. 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.

How do I find instrument spare parts when I dont know what I am looking for?

There are two ways to find instrument spare parts:

1. Using the Graphical Parts Locator:

a. Click the Services & Support tab > Find Spare Parts > and Graphical Parts Locator.

b. Select a product from one of the dropdown lists, and select Waters Quality Parts Locator.

c. 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.

2. Using the Spare Parts Catalog:

a. Click the Services & Support tab > Find Spare Parts > and Spare Parts Catalog.

b. Select a Value from one of the Menus (Product Category etc.) to begin.

c. 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.

I already know my part number

1. 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.

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:

Waters Shopping FAQs

Waters e-Commerce

How to order

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Purchasing 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 purchases.

Waters supports:

1. Catalogs via PunchOut 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 such 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 such 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, such as PunchOut (real-time pricing, availability, product pictures and specifications) or CIF (Catalog Interchange 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]

mailto:[email protected]://www.waters.com

1. Click Cart to display your shopping cart.

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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.

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How do I check out?

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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. Then 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

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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. (These 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. The 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.

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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:

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3. Click Send.

How do I see related items?

1. Click the Products tab > Chromatography Consumables & Columns > then select a Product Area.

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NOTE: Not all products have related products available.

How do I view my order history?

To access your order history, from the Waters Home Page (after login):

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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.

Common Terminology & Definitions

Not Available online

This 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.

hOW TO ORdER

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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

11

[ 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

SaMPLE PREPaRaTION

12 Sample Preparation www.waters.com

The 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 analy-sis 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.

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.

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


SaMPLE PREPaRaTION

13Sample Preparationwww.waters.com

SPE Application Area/Sorbent Selection Guide

Sorbent/SurfaceCommon

Sample MatricesDescription Typical Applications Properties

Reversed-Phase

Oasis HLBN-Vinylpyrrolidone-

DVB 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.

Drugsandmetabolitesinbiofluids

Isolationofpeptidesandoligonucleotides

High-throughputbiopolymerdesalting

Traceorganics,prioritypollutants,endocrinedisrupters

JPMHLW official food methods: antibiotics, pesticides

Particle sizes: 30 and 60 m

Pore size: 80

Surface area: 830 m2/g

pH range: 014

Sep-Pak tC18Silica

SiC18H37

Strongly hydrophobic, silica-based bonded phase used to

adsorb analytes from aqueous solutions. Trifunctional

bonding chemistry for increased hydrolytic stability.

JPMHLW official methods for pesticides in water

JPMHLW official methods for odorants in water

Particle size: 3755 m

Pore size: 125


Carbon load: 17%

pH range: 28

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


Pore size: 125


Carbon load: 12%

pH range: 28

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.

Drugsandtheirmetabolitesinbiofluids

Peptidesinserumandplasma


Pore size: 125


Carbon load: 9%

pH range: 28

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 Particle size: 3755 m

Pore size: 125


Carbon load: 2.7%

pH range: 28

Reversed- or Normal-Phase

Sep-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 exchanges in aqueous medium

below pH 8.

Phenols,phenolicpigments,naturalproducts

Petroleumfractionation

Saccharides

Drugsanddrugmetabolites

JPMHLW official methods for pesticides in food


Pore size: 125


Carbon load: 3.5%

pH range: 28

Sep-Pak CyanopropylSilica

Si(CH3)(CH2)3CN

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.

Drugsandtheirmetabolites

Pesticides


Pore size: 125


Carbon load: 6.5%

pH range: 28

Sep-Pak Diol Silica

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]


Pore size: 300


Carbon load: 2%

pH range: 28

Air Petroleum Biological Soil Water Food

Matrix Key


SaMPLE PREPaRaTION

14 Sample Preparation www.waters.com



Normal-Phase

Sep-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.

Vitaminsandfoodadditives

Lipidclassification

Syntheticorganiccompounds

Naturalproducts,plantpigments



Pore size: 125


Activity: High [ 3.2% water]

Sep-Pak FlorisilMgOSiO2

Polar, highly active, weakly basic sorbent for the

adsorption of low to moderately polar species from

non-aqueous solutions.

AOAC and EPA official methods for pesticides


Polychlorinatedbiphenyls[ PCBs]intransformeroil


Pore size: 60

Activity: High [ 2.5% water]

pH of 10% aqueous slurry: 8.5

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]


Pore size: 120

Activity: High, 1 on Brockmann

scale [ 1.5% water]

pH of 10% aqueous slurry:

A: 4 N: 7.5 B: 10

Dispersive

DisQuESpecific Application

Prepackaged centrifuge tube containing anhydrous

sodium acetate/anhydrous magnesium sulphate and

anhydrous magnesium sulphate/primaryl secondary

amine sorbent. Used for dispersive solid-phase extrac-

tion, 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-Mode

Oasis 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


Pore size: 80


pH range: 014 [pKa: < 1]

IEX capacity: 1 meq/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.

Stronglybasiccompoundsinbiofluidsandtissueextracts


and quantitation

JPMHLW official method for streptomycin and

dihydrostreptomycin in vegetable crops


Pore size: 80


pH range: 014 [pKa: ~ 5]

IEX capacity: 0.75 meq/g

Sep-Pak Accell Plus CM Acrylic 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.

Isolationofcationicproteins

Pesticides,herbicides

Steroids

Inorganiccationsinenvironmentalsamples

Particle sizes: 3755 m

Pore size: 300

pH range: 29

Ligand density: 350 moles/g

Protein-binding capacity:

175 mg Cytochrome c/g

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


and quantitation

Foodadditivesandcontaminants


Pore size: 80


pH range: 014 [pKa: > 18]


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.

Isolationofanionicproteins

Acidicpigmentsinwine,fruitjuices,foodextracts

Phenoliccompounds

Peptidepoolfractionation

Inorganicanionsinenvironmentalsamples

Particle sizes: 3755 m

Pore size: 300

pH range: 29

Ligand density: 220 moles/g

Protein-binding capacity:

200 mg BSA/g

Oasis WAX N-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


and quantitation

Emergingcontaminants


Pore size: 80


pH range: 014 [pKa: ~ 6]


Sep-Pak PSA SiC2H4NHC2H4NH2

Silica-based bonded phase containing primary and

secondary amines. Similar selectivity to aminopropyl but

with higher pKas and increased ion-exchange capacity.

Strongaffinityforfattyacids,polarpigments,andsugars

Potentialforadsorptionbychelation

Particle sizes: 37-55 m

Pore size: 60


pH range: 2-9

IEX capacity: 1.75 meg/g


SaMPLE PREPaRaTION

15Sample Preparationwww.waters.com



Products for Environmental and Food Analysis

Sep-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.

EPA Method TO11A; ASTM D5197 for carbonyl

compounds in air

JPMOE Official Methods for aldehydes: odor in out

door air and in exhaust gas


Pore size: 125

Recommended maximum capacity:

75 g [2.5 moles]

formaldehyde/cartridge

Sep-Pak XPoSure Aldehyde SamplerDiphenylhydrazine 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. Larger particle size optimized

for low-pressure personal air monitors.

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


Pore size: 125

Recommended maximum capacity:

70 g [2.3 moles]

formaldehyde/cartridge

Sep-Pak Ozone ScrubberPotassium iodide

Potassium iodide cartridge is used in series with Sep-Pak

DNPH and XPoSure Aldehyde Sampler cartridges to

remove ozone interferences.

EPA Method IP6A and ASTM D5197 for carbonyl

compounds in air

Quantity: 1.4 g KI

Capacity: 4.2 mmoles

ozone/cartridge [theoretical]

Sep-Pak DryAnhydrous Sodium Sulfate

High-capacity desiccant used to remove residual water

from normal-phase SPE extracts [in water-immiscible

organic solvents].

Generalpurpose Quantity: 2.85 g

anhydrous Na2SO4Theoretical capacity: 3.6 g H2O

Sep-Pak Porapak RDX DVBN-Vinylpyrrolidone

copolymer

Hydrophobic polymer for explosives analysis. Explosivesingroundandsurfacewateratppblevel

EPA Method 8330 for explosives


Pore size: 200

Typical sample volume:

500 mL water/cartridge

Sep-Pak PS2 StyreneDVB copolymer

Very hydrophobic copolymer designed for multi-residue

pesticide analysis in water samples.

JPMHLW official methods for pesticides in water


Particle size: 80 m

Quantity: 265 mg/cartridge

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 method for propham


[carbon black, top layer]

55105 m [aminopropyl silica]

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. Particle size: 37-105 m

[carbon-black, top layer]

37-55 m [PSA, bottom layer]

Quantity: 500 mg of each sorbent,

separated by frit

*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.

Air Petroleum Biological Soil Water Food

Matrix Key


SaMPLE PREPaRaTION

16 Oasis Solid-Phase Extraction (SPE) Products www.waters.com

Oasis Solid-Phase Extraction (SPE) Products A 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 specially-designed polymeric sorbent that performs optimally for reversed-phase SPE. The 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.

The 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 chromatographic retention (capture) of the analytes is reduced, resulting in poor recovery. Traditional, silica-based 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.

Unique Water-Wettable Oasis HLB Copolymer

N-VINYLPYRROLIDONE DIVINYLBENZENE

HYDROPHILIC-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 mm

Pore Dewetting Mechanism of Sorbent Pores (Silica Based C18)

Properly Wetted Pore Dewetted Pore If air passes through traditional C18 cartridge the pores dry out.

When sample is loaded, the analytes are not retained resulting in poor recoveries.

Analytes

not

retained

Analytes

properly

retained

in pore

Current 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


SaMPLE PREPaRaTION

17Oasis Solid-Phase Extraction (SPE) Productswww.waters.com

Oasis HLB Cartridge (30 mg) C18 Cartridge (100 mg)

% R

ecov

ery

Drying Time

ProcainamideAcetaminophenRanitidinePropranololDoxepin

Drying Time

% R

ecov

ery

0

20

40

60

80

100

0 4 8 Min.0

20

40

60

80

100

0 5 10 Min.

No impact of sorbent drying on HLBHigh, consistent recovery

Effect of Drying on RecoveryOasis HLB Versus C18 Sorbents

The 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).

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).

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 batch-to-batch reproducibility and therefore require reservations of specific lots of sorbent for large projects. Oasis sorbents have demonstrated excellent long-term, 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. The 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.

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

Higher Retention Means Greater Capacity, No Breakthrough

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

0 50 100

HO

OHO

OHNH3C

O

H

HO

HO

N

N

N

N

O

H

CH 3

CH 3

O

3X

2X

6X

15X

C18 Cartridge Oasis HLB Cartridge

Retention Factor (k) Comparison

Salicylic acid

Acetaminophen

Theobromine

Catechol

Batch-to-Batch Reproducibility of Oasis HLB Sorbent

No batch reservations needed: consistently high recoveries over more than 10 years of production

85

95

105

% R

ecov

ery

Procanamide 1.90% RSD

Ranitidine 1.68% RSD

Acetaminophen 1.65% RSD

Recovery for:

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 challenges.

Download your free Oasis Applications Notebook at www.waters.com/oasis

Oasis 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 White 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

Literature References

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.

http://www.waters.com/oasishttp://www.waters.com

SaMPLE PREPaRaTION


Oasis 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-exchange groups on the sorbent surface. T he Oasis MCX sorbent has a tightly controlled ion-exchange capacity (1 meq/g). There 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.

Oasis MCX for Basic Compounds

Drug/Sorbent Interactions on Oasis MCX Sorbent

SO3

SO3

NO

MCX sorbent

Mixed-mode Cation eXchange

Reversed-phase

interaction

Strong cation-

exchange mode

Best retention at least

2 pH units below pKaof the analyte

+CH3

CH 3

O

OH

NHH

Propranolol

(basic drug)

Sulfonic-acid-cation-

exchange capacity:

1.0 meq/g

Highly selective retention enables much stronger washes,resulting in very clean extracts.

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

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 reten-tion. If your goal is to capture basic analytes and then wash out interfer-ences 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 reversed-phase retention. We can elute with a combination of high pH and high organic concentration.

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 Mode

Base (Ionized)

Base (Unionized)

Cation Exchange

Reversed Phase

0

10

20

30

Rete

ntio

n (k

) 40

50

60

1

Reversed-PhaseRetention

Cation-ExchangeRetention

Basic Analyte(Combined)Retention

2 3 4 5 6 7

Mobile Phase pH

8 9 10 11 12 13 14

Load:Spiked and acidified sample

Condition:CH3OH

Equilibrate:H2O

Wash 1:2% HCOOH in H2O

Elute 1(Wash 2):CH3OH

Elute 2:5% NH4OH in CH3OH

Evaporate and Reconstitute:20% CH3OH in H2O

Generic Oasis MCX Method for Extraction of Basic Compounds


SaMPLE PREPaRaTION


Oasis MAX Mixed-Mode Anion eXchange and Reversed-Phase Sorbent for High Selectivity and Sensitivity for Acidic Compounds

The 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 mechanisms (anion-exchange and reversed-phase) which can be manipulated very predictably (please refer to the Oasis MAX Retention Map, below).

Oasis MAX for Acidic Compounds

Drug/Sorbent Interactions on Oasis MAX Sorbent

R+

CH2

CH3

CH3

C H4 9N

NO

MAX sorbent

Mixed-mode Anion eXchange

Reversed-phase

retention

Suprofen

(acidic drug)

Quaternary-amine-

anion-exchange capacity:

0.25 meq/g

S

CH 3O

COO

Strong anion-

exchange mode

Best retention at least

2 pH units above pKaof the analyte

+


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-Phase

0

10

20

30

40

50

60

1 2 3 4 5 6 7 8 9 10 11 12 13 14

Acid (Ionized)

Acid (Unionized)Anion Exchange

Reversed PhaseRet

entio

n (k

)

Reversed-PhaseRetention

Anion-ExchangeRetention

Acidic Analyte(Combined)Retention

Mobile Phase pH

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.

Generic Oasis MAX Method for Extraction of Acidic Compounds

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 reversed-phase retention. We can elute with a combination of low pH and high organic concentration.

Condition:CH3OH

Equilibrate:H2O

Load:Spiked acidified sample

Wash 1:5% NH4OH in H2O

Elute 1 (Wash 2):CH3OH

Elute 2:2% HCOOH in CH3OH


SaMPLE PREPaRaTION


1

0 2 4 6 8 10 Min. 0 2 4 6 8 10 Min.

2

I.S.3 4 I.S.

Results (n=6)

Drugs g/mL Type of Drug % Recovery % RSD

Nortriptyline 5 Basic (Elute 1) 94.34 0.78 Secobarbital 10 Neutral (Elute 1) 88.53 1.03 Ketoprofen 5 Acidic (Elute 2) 91.64 2.35 Naproxen 2.5 Acidic (Elute 2) 103.43 3.04

Sample

Blank with I.S.

Sample

Blank with I.S.

AV

AV

Elute 1 (basic and neutral drugs) Elute 2 (acidic drugs)

* 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.

Column: SymmetryShield 3.5 m, 4.6 x 100 mm SymmetryShield Guard column, 5 m, 3.9 x 20 mmMobile Phase: 20 mM potassium phosphate, pH 2.7/methanol, 52:48 v/vDetector: UV @ 214 nmTemperature: 30 CFlow Rate: 2 mL/minInjection Volume: 10 L of extract with internal standard

Peak Identification1. Nortriptyline2. Secobarbital3. Ketoprofen4. NaproxenI.S. Butylparaben

High Recovery of Acidic, Basic, and Neutral Compounds Using the Generic Oasis MAX Method

Oasis 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 White 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

Literature References

See Page 209

The PST Therapeutic 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. The kit contains an Oasis PST

Elution Method Development Plate, a PST 300 C18 reversed-phase column

and the detailed screening protocol.

PST Therapeutic Peptide Method Development Kit


SaMPLE PREPaRaTION


Generic Oasis WCX Method for Strong Basic Compounds

Condition:CH3OH

Equilibrate:H2O

Load:Spiked acidified sample

Wash 1:5% NH4OH in H2O

Elute 1 (Wash 2):CH3OH

Elute 2:2% HCOOH in CH3OH


The Oasis WCX (Weak Cation eXchange) SPE material was developed to provide better sample preparation for strong bases and quaternary amines. The retention mechanism is mixed mode, both ion exchange and reversed phase, which improves retention for all types of basic analytes, especially strong bases.

Oasis WCX for Strong Basic Compounds

Reversed-phase

interaction

Weak cation-exchange

interaction

WCX sorbent

Weak Cation eXchange

COO -

N

N+

O

OCH3

CH3

CH3

H C3

CH O3


Carboxylic-acid-cation-

exchange capacity: 0.75 meq/g

Excellent Recovery for Quaternary Amines as well as Polar and Hydrophobic Bases

Atenolol 10 ng/mL101% Recovery

Valethamate 10 ng/mL106% Recovery

Protriptyline 10 ng/mL102% Recovery

MRM266.9 > 144.9

MRM306.1 > 218.9

MRM264 > 191.1

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.

Column: XTerra MS C18 IS 2.1 x 20 mm, 3.5 mMobile Phase A: 10 mM NH4HCO3, pH 10Mobile Phase B: MeOH with 10 mM NH4HCO3, pH 10Gradient: 5 to 95% B in 3 minFlow Rate: 0.4 mL/minInjection Volume: 10 L

Instruments:Waters 2777 Sample Manager, and Waters 1525 binary HPLC pump, Waters Quattro Ultima

(ESI in MRM)

Recovery Data for Three Basic Compounds from Oasis WCX vs. Silica-Based WCX Products

Reco

very

(%)

120.0

100.0

80.0

60.0

40.0

20.0

0.0

Reco

very

(%)

120.0

100.0

80.0

60.0

40.0

20.0

0.0

Loading volume (mL)

0.25 1.00

Loading volume (mL)

0.25 1.00

(a)

Protriptyline (H)

Valethamate (Q)

Atenolol (P)

Protriptyline (H)

Valethamate (Q)

Atenolol (P)

Oasis WCX 96-well Elution Plate

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

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.

High Selectivity and Sensitivity for Strong Basic Compounds


SaMPLE PREPaRaTION


T he Oasis WAX (Weak Anion-eXchange) 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.

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.

SPE recoveries for three types of acidic compounds spiked in saline pH 7 and 1.3, and in rat plasma pH 1.3.

Oasis WAX for Strong Acidic Compounds

+NH

NO

+NH

Reversed-phase

interaction

Weak anion-exchange

interaction

WAX sorbent

Weak Anion eXchange

SO3

2


Piperazine-anion-exchange

capacity: 0.6 meq/g

Recovery Data for Three Acidic Compounds from Oasis WAX 1 cc, 30 mg Cartridge

40

50

60

70

80

90

100

Saline. pH = 7 Saline, pH = 1.3 pH 1.3 Rat Plasma

% R

ecov

ery 2-Naphthalenesulfonic

acid

Ketoprofen

Salicylic Acid

Recovery Data of Three Acidic Compounds for Oasis WAX vs. Silica-Based WAX Products

0

20

40

60

80

100

Oasis

WAX1 cc/

30 mg

DEA1 cc/

100 mg

PSA1 cc/

100 mg

Breakthrough at Load Step

% R

ecov

ery

Oasis

WAX1 cc/

30 mg

DEA1 cc/

100 mg

PSA1 cc/

100 mg

Recovery at Elution Step

2-Naphthalenesulfonicacid

Ketoprofen

Salicylic Acid

Spiked Saline,pH = 7

Generic Oasis WAX Method for Strong Acidic Compounds

Excellent Recovery for Strong Acid: Camphorsulfonic Acid

Concentration: 10 pg/L

108.8% recovery

Concentration: 50 pg/L

114.4% recovery

1

100

%

1.59

1.29 4.202.75 3.104.33

0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5 min0

100

%

1.59

HPLC Conditions:Column: SunFire C18 2.1 x 20 mm IS, 3.5 mMobile Phase A: H2O with 10 mM CH3COO-NH4+, pH 7Mobile Phase B: MeOH with 10 mM CH3COO-NH4+, pH 7 Flow Rate: 0.4 mL/minGradient: 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 Injection Volume: 10 LTemperature: AmbientInstrument: Waters 2777 Sample Manager, Waters 1525 Binary HPLC Pump and Waters Quattro Premier

MS/MS Conditions:Waters Quattro PremierESI- Source Temp: 150 C Desolvation Temp: 350 C Cone Gas Flow: 50 L/Hr Desolvation Gas Flow: 600 L/Hr Collision Cell: 2.2e-3 bar (Ar gas)

MRM Transition Cone (V) CID (eV) m/z 231.1 _ 79.8 60 30

(1S)-Camphor-10-sulfonic acidMW 232.3, C10H16O4S

CH3

CH2SO3H

CH3

O

High Selectivity and Sensitivity for Strong Acidic Compounds

Load:Spiked and acidified sample

Condition:CH3OH

Equilibrate:H2O

Wash 1:2% HCOOH in H2O

Elute 1(Wash 2):CH3OH

Elute 2:5% NH4OH in CH3OH



SaMPLE PREPaRaTION


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

20

40

60

% R

ecov

ery

80

100

0

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)

Acids

Neutrals

Bases

n=6RSDs < 3.5%

Ibupro

fen

Napro

xen

Salic

ylic A

cid

Sulfa

diazin

e

Sulfa

meraz

ine

Aceta

mino

phen

Theob

romine

Paraz

anthi

ne

Theop

hyllin

e

Caffe

ine

Procai

nami

de

Ranit

idine

Oxyco

done

Propra

nolol

Naltr

exone

Salbu

tamol

Doxep

in

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).

This 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.

Oasis HLB for Reversed-Phase SPE

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

Applying 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

Sample Pretreatment Suggestion

Condition:1 mL CH3OH

Equilibrate:1 mL H2O

Load:1 mL spiked sample**

Wash (Mild Wash):1 mL 5% CH3OH in H2O

Elute (Strong Elution):1 mL CH3OH

* Volumes are given for the Oasis HLB cartridge 1 cc/30 mg

Evaporate and Reconstitute:40 C/under nitrogen stream

200 L mobile phase

SO3SO3

NO

R+CH 2

CH3

CH3

C H4 9N

NO

+

Oasis HLB Sorbent

With silica-based sorbents

(C18) you would need severa

l

different chemistries to

achieve the results

shown here


SaMPLE PREPaRaTION


Neutrals WeakerBases

WeakerAcids

For Bases pKa 2-10:

Use Oasis MCX

For Acids pKa 2-8:

Use Oasis MAX

For Strong Bases pKa >10:

Use Oasis WCX

For Strong Acids pKa

SaMPLE PREPaRaTION


Oasis Sorbent Selection Plate: Evaluating Oasis 2x4 Method for Cephalexin

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:

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

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

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