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Click on Table of Contents. Then type desired page number & click OK. SAMPLE PREPARATION Solid-Phase Extraction Introduction...15 Oasis SPE Products A Breakthrough in SPE...18 Oasis HLB Cartridges, Plates, and Columns for Reversed-Phase SPE...20 Oasis MCX Cartridges, Plates, and Columns for Cation Exchange...24 Oasis MAX Cartridges, Plates, and Columns for Anion Exchange...25 Automation for Oasis SPE Products...27 High Throughput 96-well Extraction Plates...28 NEW Oasis µelution Plates...28 Oasis On-Line Columns...31 Prospekt Cartridges...33 Oasis Glass Cartridges...33 Oasis Products Selection Guides...34 Ordering Information...36 SAMPLE PREPARATION Sep-Pak Cartridges and Accessories NEW Sep-Pak 96-well Plates...39 Which Cartridge is Best for your Application...40 Which Sorbent is Best for Sep-Pak Cartridge Selection Guide...42 your Application...44 Sep-Pak Cartridges...47 Sep-Pak Vac Cartridges...48 Sep-Pak DNPH Cartridges...51 Waters XpoSure Cartridges...52 Porapak Rdx Cartridges...53 The Waters Extraction Manifold...54 13

Why SPE? Sample preparation is a key component of every analytical analysis (such as LC/MS/MS). By some estimates, 60-80% of the work activity and operating cost in the analytical laboratory is spent preparing samples for introduction into the system. The importance of sample preparation, in particular SPE, stems from three major concerns removing interferences from sample matrix, concentrating analyte(s) of interest, and improving analytical system performance. SPE offers solutions to these major concerns of sample preparation: Removing interferences from sample matrix Solid-phase extraction (SPE) has been proven to be an effective tool for selectively removing interferences, enabling sensitive, selective and robust LC/MS/MS analysis. Concentrating Analyte(s) of Interest Frequently, compounds of interest are present in levels too low for accurate and precise quantitation. SPE enables the enrichment of selected analytes without concentrating the interferences. Improving analytical system performance Advances in SPE technology combined with robotic automation make SPE not only cost effective, but also a time efficient sample preparation technique, which improves analytical system performance by: Introducing the analytes in a MS compatible solvent. Extending analytical column lifetime, reducing system downtime/maintenance. Convenient Technology for Improved Laboratory Operations Convenience and ease-of-use are primary benefits of solid-phase extraction cartridges. This can allow you to maximize the productivity of your analytical operation while saving you time and money. When compared to other sample preparation techniques, SPE cartridges also offer: Faster sample prep average time reduced by 2/3 Lower cost less solvent and reagent consumption and less hazardous waste for disposal Greater recoveries minimal sample transfer Greater accuracy no cross contamination Powerful trace concentration (enrichment) of analytes Less sample handling no emulsion problems Reduced harm to labile samples minimal evaporation Improved safety due to reduced solvent/sample exposure and glassware Easy automation simultaneous batch processing of multiple samples To satisfy all your methods development needs, Waters offers a complete range of different SPE cartridge and device designs. In addition, two different product lines are available: SEP-Pak products containing traditional silica or alumina based sorbents, and Oasis SPE products containing state of the art polymeric based sorbents. SAMPLE PREPARATION Minimizing ion suppression while improving signal response. There are many techniques used in sample preparation. Common methods include liquid-liquid extraction, centrifugation, filtration, precipitation, and now more importantly, solid-phase extraction (SPE). Traditionally, the benefits, resulting from the chromatographic technology used in solid phase extraction, were not experienced by many analysts. This was due to several factors: a lack of awareness of how to be successful with the technique, simple methods development protocols and products that did not perform reproducibly. Solid-Phase Extraction (SPE) Today, advances in SPE product technology, as well as robotic equipment make this technique capable of very cost effective and time efficient sample preparation. SPE can be one of the simplest, yet most effective and versatile, methods of sample preparation. Utilizing low cost, pre-packed, disposable cartridges containing a chromatographic packing (sorbent), a sample analyte of interest is separated from other species in the sample matrix by loading the sample onto the device and selectively eluting the desired compound(s) using different solvents. Strategies for Solid-Phase Extraction There are two simple solid-phase extraction strategies for sample preparation. You can choose a cartridge sorbent, sample solvent and elution solvent(s) to cause: Strategy 1. Compound(s) of interest are unretained, while matrix interferences are adsorbed (captured) Load Sample Matrix Waters Sep-Pak Vac Elute Compound(s) of Interest Capture (retain) Interferences then Dispose of Cartridge 15

Strategy 2. Compound(s) of interest are retained, while matrix interferences are washed from the cartridge. The compound(s) of interest are then eluted with a stronger solvent. Load Sample Matrix Elute Compound(s) of Interest with Stronger Solvent Normal Phase Chromatography (polar sorbents) with Sep-Pak Cartridges To perform normal phase chromatography with Sep-Pak cartridges, use a gradient of nonpolar solvents with polar Silica, Florisil, NH 2, Diol, CN, Alumina A, B, or N as a sorbent*. 1. You may condition the cartridge with six to ten hold-up volumes of nonpolar solvent, usually the sample solvent. Waters Sep-Pak Vac Capture (retain) Compound(s) of Interest Waters Sep-Pak Vac 2. Load the sample into the cartridge. 3. Wash unwanted compounds with a nonpolar solvent. 4. Elute the first compound of interest with a polar solvent. 5. Elute remaining compounds of interest with progressively more polar solvents. Wash out Interferences (Dispose) Collect Compound(s) of Interest as they Elute from Cartridge 6. When you recover all of your compounds, discard the used cartridge in an appropriate manner. * Depending upon your chromatographic conditions, you may also use CN as a packing material for normal phase chromatography. The first strategy is usually chosen when the desired sample compound is present in high concentration. When compounds of interest are present at low levels, or multiple compounds of widely differing polarities need to be isolated, the second strategy is generally employed. The second strategy may also be used for trace enrichment of extremely low level compounds and concentration of dilute samples. A complex matrix may be treated by both elution strategies to isolate different target analytes. Modes of Chromatography With either strategy, there are three different chromatographic modes to choose from. You can select : normal phase reversed-phase Generic Elution Protocol for Normal Phase Chromatography on Sep-Pak Cartridges (Silica, Alumina, Florisil, Diol, NH 2 ) ion-exchange The basic characteristics of each chromatographic mode are described below. There are many different types of sorbents for each mode and the Load Sample Using progressively more polar solvents to elute first and second compounds of interest 2 selection of strategy, mode, sorbent, and elution solvents will depend upon the specific sample mixture and goal of the separation. This is a general guide for the steps you would use in a method. You may condition cartridge with nonpolar solvent Elute unwanted compounds (wash) Compound 1 Compound 2 Discard the used cartridge 16

Reversed-Phase Chromatography (non-polar sorbents) with Oasis HLB and Sep-Pak Cartridges Ion-Exchange Chromatography (charged sorbent particle surface) Sep-Pak Cartridges To perform reversed-phased chromatography with Sep-Pak cartridges, use a gradient of strongly to weakly polar solvents with nonpolar C 18, tc 18, C 8, tc 2, Diol, NH 2 or CN as a sorbent. For improved performance, use the nonpolar, polymeric Oasis HLB sorbent. Oasis products provide simple, recommended methods to save time. In addition, they provide two significant benefits: increased sorbent capacity for smaller cartridge sizes, yielding higher sensitivity methods, as well as the ability to remain wetted, therefore avoiding the drying out effect of traditional sorbents that causes poor recoveries (see page 18). 1. Condition (wet/solvate) the sorbent with six to ten cartridge hold-up volumes of methanol or acetonitrile. Equilibrate the cartridge with six to ten hold-up volumes of water or buffer. (For silica based sorbents -- Do not allow the cartridge to dry out (dewet). This is not a problem with Oasis HLB devices). 2. Load the sample dissolved in strongly polar solvent. 3. Wash unwanted compounds with a strongly polar solvent. 4. Elute weakly held compounds of interest with a less polar solvent. 5. Elute more tightly bound compounds with progressively more nonpolar solvents. 6. When you recover all of your compounds, discard the used cartridge in an appropriate manner. To perform ion-exchange chromatography with Sep-Pak cartridges, use a step gradient of ph or ionic strength with Accell Plus CM, Accell Plus QMA or NH 2 as a sorbent. 1. Condition the cartridge with six to ten hold-up volumes of deionized water or weak buffer. 2. Load the sample dissolved in a solution of deionized water or buffer. 3. Elute unwanted weakly bound compounds with a weak buffer. 4. Elute the first compound of interest with a stronger buffer (change the ph or ionic strength). 5. Elute other compounds of interest with progressively stronger buffers. 6. When you recover all of your compounds, discard the used cartridge in an appropriate manner. Ion-Exchange Chromatography (charged sorbent particle surface) with Oasis MCX ( Mixed Cation exchange) or MAX (Mixed Anion exchange) sorbents. These products provide a higher level of performance and come with simple, recommended methods to save time (see pages 24 and 26). SAMPLE PREPARATION Generic Elution Protocol for Reversed Phase Chromatography on Sep-Pak Cartridges (C 18, tc 18, C 8, tc 2, Diol, NH 2, CN) Generic Elution Protocol for Ion-Exchange Chromatography on Sep-Pak Cartridges (NH 2, Accell Plus QMA, Accell Plus CM) 2 Load Sample Using progressively stronger solvents to elute first and second compounds of interest Load Sample Using progressively stronger buffers (changing ph or ionic strength, or both) to elute first and second compounds of interest Solvate the bonded phase with methanol, then flush with water and buffer Elute unwanted compounds (wash) Compound 1 Compound 2 Discard the used cartridge Condition cartridge with water or buffer Elute unwanted compound(s) with Compound 1 Compound 2 more water or buffer Discard the used cartridge For Oasis HLB recommended protocols are provided to simplify methods development even more, see page 20. For Oasis MCX and Oasis MAX sorbents ion-exchange chromatography recommended protocols are provided to simplify methods development. See pages 24 and 26. 17

Oasis SPE Products A Breakthrough in SPE Introduction In October 1977, Waters designed the first miniature cartridge columns (Sep-Pak cartridges) containing silica-based adsorbents for SPE. New demands for sample preparation led to the development of a new, specially designed polymeric sorbent which performs optimally for reversedphase SPE. The Oasis HLB copolymer with unique Hydrophilic-Lipophilic Balance is unlike traditional SPE sorbents. Today's goals for modern solid-phase extraction (SPE) are faster throughput, higher recovery and reproducibility, stronger retention and selectivity. Now SPE can outpace high throughput techniques such as LC/MS/MS. Unique Water-Wettable Oasis HLB Copolymer O N 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 Recoveries Oasis sorbents are water-wettable maintaining high retention and capacity for a wide spectrum of analytes, especially when the SPE column runs dry. When the sorbent pores dry-out, the chromatographic retention (capture) of the analytes is reduced, resulting in poor recovery. Traditional, silica based C 18 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 plate devices). Even if air passes through, the Oasis pores do not dry out. N O N O Pore Dewetting Mechanism of Sorbent Pores (Silica Based C 18 ) N-Vinylpyrrolidone Hydrophilic Lipophilic Balance Optimal Properties for Reversed-Phase SPE Specific Surface Area: 810 m 2 /g Average Pore Diameter: 80 Å Total Pore Volume: 1.3 cm 3 /g Average Particle Diameter: 5 µm, 15 µm, 25 µm, 30 µm or 60 µm* * Depending on configuration Divinylbenzene Properly Wetted Pore Analytes properly retained in pore Analytes not retained Dewetted Pore If air passes through traditional C 18 cartridge the pores dry out. When sample is loaded, the analytes are not retained resulting in poor recoveries. Current Oasis Patents: Patent No. 5,882,521, Patent No. 6,254,780, Patent No. 5,976,376, Patent No. 6,322,695, Patent No. 6,106,721, Patent No. 6,468,422, Additional Patents Pending 18

Effect of Drying on Recovery - Oasis HLB Versus C 18 Sorbents. No Stopcocks Required With Oasis Since Air Does Not Dry Out The Sorbent! % Recovery 100 80 60 40 20 Oasis HLB Cartridge (30 mg) (Excellent Recoveries) Procainamide Acetaminophen Ranitidine Propranolol Doxepin 0 0 5 10 Drying Time (minutes) 0 0 4 8 Oasis HLB 1 cc/30 mg and 1 cc/100 mg C 18 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. Higher Retention Means Greater Capacity, No Breakthrough 100 80 60 40 20 C 18 Cartridge (100 mg) (Variable Recoveries) Drying Time (minutes) The variable recoveries seen with the C 18 sorbents, due to the drying out effect, are often the cause for retests, reducing laboratory productivity. In some laboratories 10% of samples are retests this 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 C 18. Oasis sorbents work especially well when you need to capture metabolites (see figure above). High Capacity Use Less Sorbent When transferring methods from a C 18 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 C 18, This reduces breakthrough potential. In addition, you may be able to use 2 3 less sorbent than you would with C 18 (30 mg Oasis HLB gives equivalent capacity to 100 mg C 18 ). H 3 C H O HO O H N O N H N O CH 3 O CH 3 N HO N Acetaminophen OH HO Salicylic acid Theobromine Catechol Retention Factor (k) Comparison C 18 Cartridge 0 50 100 Data shown were obtained with two 3.9 mm 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 3x 2x 6x Oasis HLB Cartridge 15x Exceptional Batch-to-Batch Reproducibility Because of poor stability at ph extremes and relatively low ionic capacity traditional silica based mixed-mode sorbents don t have long-term batch-tobatch 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 6 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. Batch-to-Batch Reproducibility of Oasis HLB Sorbent Recovery (%) 105 95 85 75 65 No Batch Reservations Needed Overall Guideline for Sorbent Selection There are three available Oasis sorbent chemistries which are designed to solve just about all of your Methods Development problems. They are all built upon this unique co-polymer and provide exceptional results. The sulfonated and Quaternary amine derivatives Oasis MCX and MAX are designed for selective retention of basic, and acidic compounds respectively. Oasis HLB (Hydrophilic-Lipophilic Balance) reversed-phase sorbent. Oasis MCX (Mixed-mode: Cation-eXchange and reversed-phase) sorbent. Oasis MAX (Mixed-mode: Anion-eXchange and reversedphase) sorbent. A fast approach to successful methods development with Oasis sorbents is as follows; First, choose the best sorbent for your analyte(s), as diagramed below; Determine Types of Analyte(s) in Sample Matrix (Acids, Bases, Neutrals) If a mix of Acids, Bases and Neutrals, or Unknowns Reversed-Phase Start with Oasis HLB Recommended 1-D Method Procanamide (1.90% RSD) Ranitidine (1.68% RSD) Acetaminophen (1.65% RSD) Batch Number If all are Bases Cation Exchange Start with Oasis MCX Recommended Method Start with the Recommended Method for that sorbent. If all are Acids Anion Exchange Start with Oasis MAX Recommended Method See page 20 See page 24 See page 26 SAMPLE PREPARATION 19

Oasis HLB Plates, Cartridges and Columns This generic, 1-D method (1-Dimensional only the organic strength is changed) has proven useful for a wide variety of compound types may be the only protocol required, reducing method development time. Recommended Generic Oasis HLB SPE Method (1-D)* 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 <5%) with high recovery (>85%), using the same simple SPE protocol (see below). Condition 1 ml methanol Equilibrate 1 ml water Load 1 ml spiked sample** Wash (Mild Wash) 1 ml 5% methanol in water Elute (Strong Elution) 1 ml methanol Evaporate & Reconstitute 40 C/under nitrogen stream 200 µl mobile phase * Volumes are given for the Oasis HLB cartridge 1 cc/30 mg Sample Pretreatment Suggestion 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 High ph Low Low 2 5% 1 Organic Concentration 100% 1: Load (sample in acid disrupt binding)** 2: Wash 5% MeOH 3: Elute 100% MeOH **Add 20 µl (H 2 PO 4 ) to disrupt drug-protein interaction/ binding Dilute 1:1 or greater with 0.1 N HCl or other acids Filter through 0.45 µm membrane Centrifuge @ 3000 rpm 3 High One Simple Procedure: Many Applications on a Universal Sorbent Reduced Methods Development Time 100 % Recovery Acids Neutrals With silica-based sorbents (C 18 ) you would need several different chemistries to achieve the results shown here Ibuprofen Naproxen Salicylic Acid Sulfadiazine Sulfamerazine Acetaminophen Theobromine Parazanthine Theophylline Caffeine Procainamide Ranitidine Oxycodone Propranolol Naltrexone Salbutamol Doxepin 80 60 40 20 0 Bases n=6 RSDs < 3.5% 1-D Method Seventeen 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 µl/ml H 2 PO 4 was used to disrupt drug-protein binding). 20

Sorbent Amount and Solvent Selection for the Generic Method The suggested amount of sorbent in a cartridge or a plate required for your application, is given in the table below. Remember, because of the increased capacity of the Oasis Sorbents, you can use less sorbent than you would normally need if you used a silica based packing. The solvent used for the elution step should be selected based on the polarity of the analyte. The second table below gives a selection of elution solvents and each solvent gives you different selectivity and elution strength. Capacity and Elution Volume of Oasis 96-Well Plates and Cartridges Sorbent Maximum Typical Elution per device Mass Capacity Sample Volumes Volume µelution Plate* 60 to 400 µg 10 to 375 µl 25 µl** 5 mg* 0.15 to 1 mg 10 to 100 µl 150 µl 10 mg 0.35 to 2 mg 50 to 200 µl 250 µl 30 mg 1 to 5 mg 100 µl to 1mL 400 µl 60 mg 2 to 10 mg 200 µl to 2 ml 800 µl * Available only in 96-well plates ** µelution Plate requires no evaporation step When converting from C 18 silica-based to Oasis SPE sorbents, use approximately 2 3 less Oasis sorbent (100 mg C 18 sorbent = 30 mg Oasis sorbent). Note: Larger capacity cartridges are also available. Tips for Selecting Elution Solvents for the Generic SPE Method (1-D)* The elution solvent is selected based on polarity of analyte. Solvent Solvent Type Relative Comments Elution Strength** Methanol proton donor 1.0 disrupts H-bonding Acetonitrile dipole-dipole 3.1 medium polarity drugs Tetrahydrofuran dipole-dipole 3.7 medium polarity drugs Acetone dipole-dipole 8.8 medium polarity drugs Ethyl Acetate dipole-dipole high nonpolar drugs and GC compatible Methylene Chloride dipole-dipole high nonpolar drugs and GC compatible * When using solvents other than methanol, add 10-30%(of proton donor solvent like methanol) to disrupt H-bonding on the Oasis HLB sorbent. ** High-Purity Solvent Guide. Burdick & Jackson Laboratories, Inc. Solvent Properties of Common Liquids, L.R. Snyder, J. Chromatogr., 92, 223 (1974); J. Chromatogr. Sci. 16, 223 (1978) More Selective and Sensitive Reversed-phase Methods Development Using 2-Dimensional (2-D) Strategy for Oasis HLB Sorbent Meeting even more difficult Selectivity and Sensitivity Goals The generic 1-D method can be expanded to meet even more challenging selectivity and sensitivity goals. Using a chromatographically based approach, more powerful methods can be developed, very predictably, to remove plasma, urine or any other sample matrix interferences. A comparison of the 1-D and 2-D strategies (below) shows that you can tailor the cleanliness of the baseline depending on your needs. A cleaner extract is achieved by simply manipulating the organic concentration and the ph. This is a 2-Dimensional approach, which now includes the powerful selectivity of ph adjustment to create a more sensitive and selective method. The impact of ph for reversed-phase applications is shown on the accompanying Retention Map. This is the basis of how this method development strategy was created (see next page). Better Results with a 2-Dimensional Method Column: SymmetryShield RP 8, 5 µm, 3.9 x 150 mm Temperature: 30 C Mobile Phase: 50 mm Phosphate ph 7: acetonitrile:methanol (41:37:22); Alliance 2690 Detection: UV at 230 nm Flow Rate: 1.0 ml/min. Injection Volume: 40 µl (after evaporation and reconstitution in 200 µl water) AV AV 1-D Method (Generic) 0.004 AU 2-D Method with 2 washes 0.004 AU 2-D Method with 3 washes 1 1 Sample Blank Sample 2 2 3 3 Peak Identification: Peak 1: Norverapamil Peak 2: Verapamil Peak 3: Methoxyverapamil (I.S.) High ph Low Low 1: Load 2: Wash 5% MeOH 3: Elute 100% MeOH Blank High ph Low Low 1 5% 2 2 Organic Concentration 3 65% 100% 5% 1 4 Organic Concentration 1: Load 2: Wash 5% MeOH 3: Wash High ph 65% MeOH 4: Elute Low ph 65% MeOH 100% 3 High High SAMPLE PREPARATION 1 2 High 3 4 The generic method (1-D) is an excellent starting protocol for methods development. If you were not able to meet all of your SPE goals, then the advanced 2-D protocol will provide a rapid way to chromatographically determine an even cleaner, more selective and sensitive result. AV 0 0.004 AU 2 4 Sample Blank 6 8 Minutes 3 10 65% 100% ph 5% 1 2 5 Low Low Organic High Concentration 1: Load 2: Wash Low ph 5% MeOH 3: Wash High ph 5% MeOH 4: Wash High ph 65% MeOH 5: Elute Low ph 65% MeOH 21

This is called a Retention Map for Reversed-phase chromatography. We have plotted the retention (k) of a neutral, acidic and basic compound as a function of the ph. For a basic compound, it indicates that when a basic compound is ionized at low ph (2 units below its pk a ), it is poorly retained by SPE. At high ph (2 units above its pk a ), it is un-ionized, and strongly retained by reversed-phase SPE, with a high k value. This means that if we want to capture a base on our reversed-phase chromatographic sorbent, we need to LOAD and Wash at High ph, and then we can ELUTE the base in a cleaner extract at Low ph. In general, when the ionizable compound is in its unionized state you will obtain a high k, excellent capture. In its ionized state, there is very little k, therefore it will be easy to elute from the sorbent. Retention Factor Versus ph for Acids, Bases and Neutrals Retention Map, Reversed-Phase, Oasis HLB 60 Acid (Unionized) Base (Unionized) An acidic compound behaves the same way relative to its ionization. Note that the retention of a neutral compound is not changed by adjusting the ph, it is affected by the organic concentration only. Un-ionized (Captured) ph 2 ph 12 HA N BH+ Load and Wash at Low ph to capture Acid, release bases, neutrals Ionized A- Elute at High ph to release Acid Retention (k) 50 40 30 20 10 0 Base (Ionized) Neutral Acid (Ionized) 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Mobile Phase ph Recommended ph range of Silica Using this chromatographically based approach, a Wash and Elution Study is performed to determine the optimal SPE method using organic concentration and ph. (Always test your analytes for stability and solubility at different ph values.) Investing approximately 4 hours of development time, using this strategy, yields extremely powerful, and predictable results. Overall methods development time will decrease, with the added benefit of very clean, selective extracts. (This approach has solved many methods development problems over the last several years. Contact your Waters Technical Support Team for more information.) Impact of ph Recommended ph range of Oasis Base Neutral Acid ph 12 ph 2 The 2-D experiment (following page) demonstrates this approach. Twenty samples of the analytes in a saline solution (PBS) are loaded onto twenty conditioned and equilibrated cartridges or wells of a 96-well plate. Each cartridge is eluted with a different bottle of solvent (10 bottles contain Low ph solutions-each with different concentrations of methanol, and 10 High ph solutions). The eluate is then analyzed and the area count for each analyte is plotted relative to its elution bottle of solvent on two area count plots (Low ph and High ph). The area counts will be zero if the analyte is still captured on the cartridge. Un-ionized (Captured) B Reviewing the two Area Count Plots, note that the analytes begin to elute with lower organic strength (~35%) at Low ph. At High ph it takes over 70% organic. (Based on the Retention Map for Reversed-Phase, this indicates that these analytes were bases, since they were unionized at High ph, had a higher k, and required more organic for elution.) N A- Load and Wash at High ph to capture Base, release acids, neutrals Ionized BH+ Elute at Low ph to release Base From these plots, we can design a 2-D method which will greatly reduce matrix interferences in two ways relative to the generic 1-D method; first, an additional stronger, more aggressive Wash step to get rid of more interferences; and second, a weaker, less aggressive Elution step that will release our analytes, but leave other interferences captured in the cartridge. Since this is determined chromatographically, it is a very predictable approach. 22

2-D Wash Elute Study: Experiment for Oasis HLB 30 mg 96-well Plate or Cartridge 200000 150000 100000 50000 0% 10% 20% 30% 40% 50% 60% 70% 80% 90% % MeOH 0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 2-D Wash Elute Study Results (Area Counts) Base Modified Wash High ph Area (µv*sec) Prepare analytes in saline Condition/equilibrate 20 cartridges 1 ml methanol/1 ml water Load 1 ml spiked saline solution Elute with 1 ml 2% NH 4 OH in MeOH/water 2% CH 3 COOH in MeOH/water Maximum % Methanol Wash 150000 100000 50000 Acid Modified Wash Low ph Area (µv*sec) Minimum % Methanol Elution To obtain complete elution, the elution solvent should be acidic (2% acetic acid) and the methanol concentration should be greater than 60%, but less than 70%. Remember, the elution solvent should be able to fully elute the analytes, but not be too strong and contain too much organic to release undesirable neutral interferences from the sorbent. For this experiment, low ph with no more than 70% methanol should be used. Notice, we did not choose high ph for an elution condition, since this made the analytes retain more, and thus would require almost 100% methanol to elute them. This high percentage of an organic concentration would also elute undesirable interferences along with the analytes. The cleanest extract was achieved for verapamil and its metabolite using the 2-D, 3-wash procedure instead of the 1-D generic method. The more selective method enables higher sensitivity for analyses at ng/ml levels. Important Note: Traditional, Silica based C 18 sorbents should not be exposed to High ph because the silica particle substrate dissolves, which can contaminate your extracts with silicates. Oasis HLB works well because it is a copolymer, which doesn t dissolve at high ph. 2-D SPE 3-Wash Method Developed from Wash Elute Study Oasis HLB Condition 1 ml methanol SAMPLE PREPARATION 0 0 20 40 60 80 100 0 0 20 40 60 80 100 % MeOH % MeOH Equilibrate 1 ml water Acid 2% Acetic Acid Peak Area (µvs) Peak Area (µvs) Base 2% Ammonium Methoxy- Hydroxide Verapamil Norverapamil verapamil 0-70% No Response No Response No Response 80% 34,095 47,482 6,358 90% 63,158 85,704 178,178 Methoxy- Verapamil Norverapamil verapamil 0-30% No Response No Response No Response 40% 73,146 32,269 86,789 50% 121,820 68,259 137,899 60% 109,145 61,830 123,229 From these results, the wash solvent(s) can be either acidic, up to 30% methanol containing 2% acetic acid, or basic, up to ~70% methanol containing 2% ammonium hydroxide, since no analyte is eluted under these conditions (your choice depending on the type of sample matrix interferences present in some cases, you can add multiple wash steps for even more sensitive methods). If we just wanted one more wash, it should be aggressive as possible, therefore we should chose high ph and 65% methanol. Load 1 ml spiked sample Wash 1 1 ml 5% MeOH in water with 2% acetic acid Wash 2 1 ml 5% MeOH in water with 2% ammonium hydroxide Wash 3 1 ml 65% MeOH in water with 2% ammonium hydroxide Elute 1 ml 65% MeOH in water with 2% acetic acid Evaporate & Reconstitute 40 C/under nitrogen stream 200 µl water/mobile phase High ph Low Low 5% 3 4 1 2 5 Organic Concentration 65% 100% High Removes neutral polar and basic polar interferences Removes acidic polar interferences Removes neutral non-polar and basic lipophilic interferences but not analytes Elute the basic analyte compounds in ionized form 1: Load 2: Wash Low ph 5% MeOH 3: Wash High ph 5% MeOH 4: Wash High ph 65% MeOH 5: Elute Low ph 65% MeOH 23

Oasis MCX Plates, Cartridges and Columns 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, 60 Base (Ionized) 50 Retention (k) 40 30 20 Cation Exchange Reversed Phase Base (Unionized) 10 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Mobile Phase ph High Selectivity and Sensitivity for Basic Compounds Oasis MCX Sorbent for Cation Exchange Obtain selective retention of basic drugs with cation-exchange groups on the sorbent surface. The Oasis MCX sorbent has a tightly controlled ionexchange capacity (1 meq/gram). There are no silanol groups to complicate the retention mode or method development. This novel, water-wettable, polymeric sorbent is stable from ph 0 to 14, making method development simple and fast. Drug Sorbent Interactions on Oasis MCX Sorbent Reversed-Phase Retention Cation-Exchange Retention Basic Analyte (Combined) Retention This 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. MCX sorbent Mixed Mode Cation exchanger O N 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- SO 3 SO 3 phase retention. We can elute with a combination of high ph and high organic concentration. Strong cation-exchange mode best retention at least 2 ph units below pka Reversed-phase interaction CH 3 O + N H H CH 3 Sulfonic Acid Cation OH Exchange Capacity: Propranolol Basic Drug 1.0 meq/g Highly selective retention, enabling 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 retentions mechanisms (cation exchange and reversed-phase) which can be manipulated very predictably (please refer to the Oasis MCX Retention Map). Generic Oasis MCX Method for Extraction of Basic Compounds Condition 1 ml MeOH Equilibrate 1 ml H 2 O Load 3 ml spiked and acidified urine Wash 1 2 ml 0.1N HCI Wash 2 2 ml methanol Elute Bases 2 ml 5% ammonium hydroxide in methanol Evaporate and Reconstitute 300 µl 20% methanol in water Removes polar and acidic interferences Removes neutrals Oasis MCX 3 cc/60 mg cartridge 24

As shown, one protocol with minimum wash steps gives extractions fast enough to keep pace with your analytical system. In some cases, with urine samples, no conditioning or equilibration of cartridges is required to achieve excellent results for basic drugs. Oasis MAX Plates, Cartridges and Columns Comparison of Oasis MCX Cartridges to other SPE Silica-Based Products Using the Generic Oasis MCX Method 100 % Recovery (% RSD) 80 60 40 20 0 0.3% 0.7% 0.5% Oasis MCX 60 mg/3 cc 2.4% 3.2% 3.2% Brand A 300 mg/ 3 cc 13% 12% 1.1% Brand B 300 mg/ 3 cc Methadone Methadone metabolite (EDDP) Propranolol The use of the 2-D approach can be very successful for the Oasis MCX sorbent as well. A ph modified wash system allows optimization of the method and it produces a very selective protocol for basic compounds. Selectively Separate Primary and Secondary Amines from Tertiary and Aromatic Amines Using an Oasis MCX Cartridge Oasis MAX plates, cartridges and columns High Selectivity and Sensitivity for Acidic Compounds using Anion Exchange The Oasis MAX sorbent has a tightly controlled ion exchange capacity of 0.3 meq/gram ensuring reproducible SPE protocols for extraction of acidic compounds and metabolites from biological fluids. There are no silanol groups to complicate the retention mode or method development. This 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 retentions mechanisms (anion exchange and reversed-phase) which can be manipulated very predictably (please refer to the Oasis MAX Retention Map, page 26). SAMPLE PREPARATION % Recovery 100 85 75 50 25 15 Elute 1 Tertiary and aromatic amines with 5% TEA Elute 2 Primary and secondary amines with 5% NH 4 OH in methanol 0 0 200 400 600 800 1000 Elution Volume (µl of 5% TEA in Methanol) Procainamide (3 ) Toluidine (Aromatic) Doxepin (3 ) Amitriptyline (3 ) Nortriptyline (2 ) N,N-dimethylbenzylamine (3 ) N-methylbenzylamine (2 ) Benzylamine (1 ) Amphetamine (1 ) Methamphetamine (2 ) We can also fractionate the tertiary and aromatic amines in an Elute 1 step using 5% TEA (triethyl amine) in MeOH, with very little release of the primary and secondary amines. To elute the primary and secondary amines ammonium hydroxide is used. Drug Sorbent Interactions on Oasis MAX Sorbent Reversed-phase Retention R+ Quaternary Amine Anion Exchange Capacity: 0.3 meq/g S O O N MAX sorbent Mixed Mode Anion exchanger COO CH 3 CH 3 + CH 2 N C 4 H 9 CH 3 Strong anionexchange mode Best retention at least 2 ph units above pka Suprofen Acidic Drug 25

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 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. 60 Acid (Ionized) Generic Oasis MAX Method for Extraction of Acidic Drugs Retention (k) 50 40 30 20 10 0 Acid (Unionized) Anion Exchange Reversed Phase 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Mobile Phase ph Reversed-Phase Retention Anion-Exchange Retention Acidic Analyte (Combined) Retention Condition 1 ml methanol Equilibrate 1 ml water Load* 3 ml sample Wash 2 ml 50 mm sodium acetate ph 7/5% methanol Wash 2 (Elute 1) 2 ml methanol Removes/elutes basic and neutral interferences The Retention Map plots the total k or capacity (retention) of a 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. At low ph, the ion exchange retention mechanism shuts-off because the analyte becomes un-ionized. Only reversed-phase retention is present, but Elute 2 2 ml 100 mm phosphoric acid/acetonitrile 1:1 (v/v) Alternative LC/MS Elution System: 2% formic acid in methanol As shown below, acidic compounds can be separated from basic and neutral compounds on the same Oasis MAX cartridge following the recommended protocol. High Recovery of Acidic, Basic and Neutral Compounds Using the Generic Oasis MAX Method Elute 1 (basic and neutral drugs) Elute 2 (acidic drugs) 1 2 I.S. 3 4 I.S. AV AV 0 2 4 6 8 10 0 2 4 6 8 10 Minutes Column: SymmetryShield 3.5 µm, 4.6 mm x 100 mm SymmetryShield Guard column, 5 µm, 3.9 mm x 20 mm Mobile Phase: 20 mm potassium phosphate, ph 2.7/methanol, 52:48 v/v Detector: UV @ 214 nm Temperature: 30 C Flow Rate: 2 ml/min Injection Volume: 10 µl of extract with internal standard Peak Identification 1. Nortriptyline 2. Secobarbital 3. Ketoprofen 4. Naproxen I.S. Butylparaben Sample Blank with I.S. Minutes Sample Blank with I.S. Results (n=6) Drugs µg/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 *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. 26

Oasis SPE Products Oasis products come in a full range of device formats to meet your SPE requirements the new µelution plates, on-line columns, 96-well plates, and single-use cartridges. Try Oasis and successfully meet your SPE challenges. Automation Request your free Oasis Applications Notebook (Literature Code 720000609EN) at /oasis or contact your local sales office. Automation of Oasis Sample Extraction Cartridges and 96-Well Plates SAMPLE PREPARATION Oasis SPE products are compatible with the following liquid handling and/or SPE automation systems Packard MultiProbe II Robotic liquid handling system Tomtec Quadra96 and Quadra96 SV Pipetting Station Hamilton Microlab SPE Workstation Beckman Biomek 2000 Laboratory Automation Workstation Zymark RapidTrace Automated SPE Workstation Gilson ASPEC XL4 Gilson ASPEC XL Gilson 215 SPE Liquid Handler Tecan Genesis Prospekt /Prospekt II Waters LC/MS/MS Systems for Bioanalysis Featuring the Waters Quattro Premier For high sensitivity analyses, such as those employing LC/MS/MS, proper sample preparation can be critical for minimizing matrix effects and concentrating analytes of interest. Oasis Sample Extraction Products can be used with Waters LC/MS/MS Systems including this system that integrates the Waters Quattro Premier Mass Spectrometer with the Waters 1525µ Binary Pump and the 2777 Sample Manager using MassLynx 4.0 Software. 27

High Throughput 96-Well Extraction Plates The Oasis sorbents come in several different mass amounts, in a 96-well plate configuration that can be used on many manifolds and automated instruments. Because of the unique sorbent properties (no drying-out effect), each well gives high and reproducible recoveries for a wide range of analytes including polar and basic drugs, with RSDs 5% (n=96). Waters offers the standard high-throughput 30 mg and 60 mg Oasis 96-well plates as well as 5 mg and 10 mg 96-well low elution plates. Waters 96-Well Plate Design 1999 R&D 100 Award Two-Stage Well Design 5 mg 10 mg 30 mg 60 mg By varying frit size and/or placement, the same plate may be filled with various quantities of sorbent per well. Our design permits optimal recoveries, even with low sorbent weights for smaller elution volumes. Oasis µelution Plates for Ultra Low Elution Volumes Elute in as little as 25 µl with no evaporation/reconstitution Ideal for small sample volumes Up to a 25x increase in sensitivity Waters Newest Innovation in SPE Technology Scientists in both drug discovery and drug development can prepare biological samples for LC/MS/MS analysis using a more efficient, generic methodology and can successfully eliminate the time consuming evaporation step with the new Oasis µelution plate. * Patent Pending Excellent Recovery in 25 µl Elution OASIS µelution Technology >85% Recovery in 25 µl 25 µl 100 80 100 µl SPIKED SALINE acetaminophen practolol The new Oasis µelution plate combines patented* plate design, proven Oasis chemistries, and recommended protocols enabling elution volumes as low as 25 µl. Now for the first time you can perform SPE clean-up and concentration of very small sample volumes. Achieve superior results compared to protein precipitation in the same or less time using the Oasis µelution Plate. This plate produces extracts that can be directly injected, eliminating the time-consuming evaporation step. Just elute and shoot. Eluting in 25 µl without evaporation provides up to 25x increase in sample concentration, enabling sensitive, robust, and reproducible SPE results. % Recovery 60 40 20 0 Oasis HLB µelution plate N-acetyl procainamide betamethsone caffeine naproxen amitripyline propranolol The innovative features of the new Oasis µelution plates enable sensitive, robust, reproducible results without evaporation and reconstitution. 28

Speed and Throughput Comparison of Oasis µelution Plate and Protein Precipitation Superior Results, Less Time and Effort The Oasis µelution plate optimizes the configuration of the highly efficient Oasis sorbents (HLB, MCX, MAX), enabling elution volumes as low as 25 µl, providing fast clean up with improved performance over protein precipitation. Time consuming evaporation and reconstitution steps are eliminated, compressing preparation cycle time and increasing throughput capabilities. Extraction Protocols The protein precipitation generic method includes both a centrifugation and an evaporation step, which produces the cleanest sample extract possible for protein precipitation. The final sample volume (75 µl) is the same for all three generic methods. The Oasis generic methods produce cleaner, extracts than protein precipitation, demonstrated by the improved sensitivity with the generic Oasis HLB (4x) and the generic Oasis MCX (9x to 25x) methods. The Oasis methods enable improved sensitivity by reducing ion suppression. Achieve superior results compared to protein precipitation in less time using the Oasis µelution Plate. Protein Precipitation Generic Method Add 50 µl spiked plasma (1 to 1000 pg/µl), 50 µl IS (10 pg/µl) in H 2 O Add 1 ml ACN to precipitate plasma Centrifuge the plasma at 3600 rpm for 30 minutes* Transfer the supernatant to another 2 ml 96-well container Evaporate the solution to dryness* Reconstitute with 25 µl ACN:IPA (40:60) + 2% NH 4 OH and 50 µl H 2 O Inject 20 µl Oasis HLB Generic Method Oasis HLB µelution Plate** Part Number186001828 Condition with 200 µl MeOH Equilibrate with: 200 µl H 2 O Load 50 µl spiked rat plasma, 50 µl IS (10 pg/µl) in H 2 O Wash with 200 µl 5% MeOH in water Elute with 25 µl ACN:IPA 40:60 + 2% FA Dilute with 50 µl H 2 O Inject 20 µl Oasis MCX Generic Method Oasis MCX µelution Plate** Part Number 186001830 Condition with 200 µl MeOH Equilibrate with 200 µl H 2 O Load 50 µl spiked rat plasma, 50 µl IS (10 pg/µl) in H 2 O Wash 1: 200 µl Water + 2% FA Wash 2: 200 µl MeOH Elute with 25 µl ACN:IPA 40:60 + 2% NH 4 OH Dilute with 50 µl H 2 O Inject 20 µl SAMPLE PREPARATION Increase in Selectivity (More Selective Methods) ** Tomtec Quadra 96 used. Oasis HLB and MCX µelution Plate versus Protein Precipitation LC Conditions Column: XTerra MS C 18, 2.1 x 30 mm, 3.5 µm Mobile phase A: Water + 0.5 % NH 4 OH Mobile phase B: ACN + 0.5 % NH 4 OH Flow rate: 0.2 ml/min LC conditions: 5% - 95% B 1 min. Temperature: room temp. LC Instrument: Waters Alliance 2795 MS Conditions MS Instrument: Micromass Quattro Triple Quadrupole Ion source: ESI (+) Source temperature: 150 C Gas cell: 2.0 e-3 bar Argon Desolvation temperature: 350 C Drying gas flow: 500 L/hr Cone gas flow: 50 L/hr Cone voltage: 25 volts Collision energy: 20 Capillary voltage: 3.5 Kv MRM transition: Metoclopramide (IS) m/z 299.8 226.7 Propranolol m/z 259.9 154.9 Amitriptyline m/z 278.1 232.9 Nortriptyline m/z 263.9 190.8 Propranolol, 1 ng/ml Amitriptyline, 0.1 ng/ml 100 % Protein Precipitation Intensity: 7.50 x10 3 3.79 100 % Protein Precipitation Intensity: 9.71 x10 3 4.75 5.18 16 17 100 % Oasis HLB Intensity: 3.03 x10 4 3.83 4x sensitivity without evaporation 100 % Oasis HLB Intensity: 3.62 x10 4 4.67 4x sensitivity without evaporation 7 4 100 % Oasis MCX Intensity: 1.90 x10 5 3.83 25x sensitivity without evaporation 100 % Oasis MCX Intensity: 8.91 x10 4 4.48 9x sensitivity without evaporation 2 0.50 1.00 1.50 2.00 2.50 3.00 Time (mins) 3.50 4.00 4.50 5.00 2 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 Time (mins) 5.00 5.50 6.00 29

Sensitivity and Precision Comparison of Oasis MCX µelution Plate and Glass Fiber and Membrane 96-well Disk Plates Up to 4x Increase in Sensitivity The Oasis µelution plate shows superior performance when compared to other low elution disk plate products used according to recommended protocols. A generic Oasis MCX method for terfenadine with 50 µl elution, dilution and direct injection shows better sensitivity than the membrane and glass fiber plates, which require a 150 µl elution volume for complete analyte recovery. The Oasis µelution plate enables sensitivity gains and delivers high performance without the time consuming evaporation step. Oasis MCX µelution Plate versus Glass Fiber and Membrane 96-well Disk Plates Glass Fiber SCX, 96-well Plate 250 µl plasma loaded 150 µl elution Membrane MPC 96-well Plate 250 µl plasma loaded 150 µl elution Oasis MCX µelution Plate 250 µl plasma loaded 50 µl elution Terfenadine 0.5 ng/ml 100 % S/N = 61 3.02 2.92 100 2.92 100 Intensity: S/N = 82 Intensity: Intensity: 5.75 x10 6 7.44 x10 6 S/N = 357 1.14 x10 7 % 6x Sensitivity % -10 2.00 2.50 3.00 3.50-10 2.00 2.50 3.00 3.50-10 2.00 2.50 3.00 3.50 Terfenadinealcohol 0.5 ng/ml 100 % 2.54 S/N = 11 2.47 2.45 100 100 Intensity: S/N = 17 Intensity: Intensity: 3.03 x10 4 7.14 x10 S/N = 178 4 3.11 x10 5 % 16x Sensitivity % -10 2.00 2.50 3.00 3.50-10 2.00 2.50 3.00 3.50-10 2.00 2.50 3.00 3.50 Terfenadinecarboxylate 0.5 ng/ml 100 % 2.70 S/2 = 51 2.62 100 2.63 100 Intensity: S/N = 40 Intensity: Intensity: 7.30 x10 4 1.15 x10 5 S/N = 927 5.04 x10 5 % 18x Sensitivity % -10 2.00 2.50 3.00 3.50-10 2.00 2.50 3.00 3.50-10 2.00 2.50 3.00 3.50 Time (mins) Time (mins) Time (mins) Final extract diluted 1:2 with H 2 O Extracted using manufacturers recommended protocol Final extract diluted 1:2 with H 2 O Extracted using manufacturers recommended protocol Final extract diluted 1:2 with H 2 O Extracted using manufacturers recommended protocol Validation Assay of Terfenadine and Its Metabolites Using The Oasis MCX µelution Plate Calibration range: 0.5-200 ng/ml, 7 levels 6 Calibration curves per day over four days Analyte recoveries over calibration range: 95% Inter-day %RSD of back-calculated standard concentrations over four days (24 values) 5.6% Intra-day %RSD of back-calculated standard concentrations within one day (6 values) 8.0% 30

Oasis On-Line Columns for LC/MS or LC/MS/MS Generally, drugs present in biological matrices should not be directly introduced into the LC/MS/MS. If they are directly injected onto the system, column clogging and analyte ion suppression due to the presence of proteins, lipids, salts and other endogenous material are inevitable. Therefore a sample cleanup is a must before LC/MS/MS analysis. In addition to the off-line sample cleanup, on-line sample preparation also provides a fast and efficient way to cleanup the sample before LC/MS/MS analysis. In the Oasis On-Line Column for LC/MS/MS application, a diluted plasma sample is injected onto the Oasis On-Line column. The column is then purged of most protein and interferences. A fast gradient is run to elute the target analyte, a fraction of this flow is diverted to the mass spectrometer for detection (see next page). Oasis Patented Chemistry for Oasis On-Line Columns There are three available Oasis sorbent chemistries which are designed to meet just about all of your on-line LC/MS/MS needs. They are all built upon unique water-wettable Oasis HLB copolymer and provide exceptional results. The sulfonated and Quantenary amine derivatives Oasis MCX and MAX are designed for selective retention of basic and acidic compounds respectively: Oasis HLB (Hydrophilic-Lipophilic Balance) reversed-phase sorbent. Oasis MCX (Mixed-mode: Cation-eXchange and reversed-phase) sorbent. Oasis MAX (Mixed-mode: Anion-eXchange and reversed-phase) sorbent. Oasis sorbents are water-wettable enabling high capacity and retention of a wide spectrum of analytes. This feature of the Oasis sorbent is especially important when the Oasis On-Line column is cycled from 100% aqueous to 100% organic mobile phase. Also the polymeric nature of Oasis enables column use under ph extremes (ph 1-14). (Note: low and variable results are often obtained for silica-based columns under such conditions) Oasis On-Line columns also provide exceptional batch-to-batch and column-to-column reproducibility. Unique Water-Wettable Oasis HLB Copolymer N O N-Vinylpyrrolidone O N Hydrophilic Lipophilic Balance Optimal Properties for Reversed-Phase SPE Specific Surface Area: 810 m 2 /g Average Pore Diameter: 80 Å Total Pore Volume: 1.3 cm 3 /g Average Particle Diameter: 5 µm, 15 µm, 25 µm, 30 µm or 60 µm* * Depending on configuration N O Divinylbenzene SAMPLE PREPARATION O N O N SO 3 SO 3 R+ CH 3 + CH 2 N C 4 H 9 CH 3 Oasis HLB Sorbent Oasis MCX Sorbent Oasis MAX Sorbent 31

Oasis On-Line Columns Ultra fast On-Line SPE for LC/MS/MS Ultra fast on-line analysis can improve the speed and throughput of LC/MS analysis of small molecules from biologic sample matrices.there are three Oasis On-Line column configurations designed to fit all your on-line analyses needs. The Oasis cartridge column fits into a Sentry holder that features a finger-tighten fitting for fast and convenient replacement (1). The direct connect column can be screwed directly into a switching valve or connected to fittings like a conventional HPLC column (2). The Oasis column features traditional HPLC column fittings and hardware (3). 1 3 2 All of these formats are available with the three Oasis patented sorbents (HLB, MCX, and MAX), and in a wide choice of particle sizes as well as dimensions. The Oasis On-Line columns make it possible to analyze a specific analyte or metabolite from a complex sample matrix (such as plasma or serum). No off-line sample preparation is needed; just dilute the sample and inject. Ultra fast on-line analysis of multiple analytes can also be accomplished using the Oasis On-Line column combined with Waters narrow-bore analytical columns (such as Atlantis, XTerra, or Symmetry columns). Ultra Fast On-Line System Configuration Oasis On-line Column LOAD POSITION 515 System INJECTION POSITION 515 System Isocratic HPLC Waste 1 10 9 1 10 9 10 Port Valve 2 positions Waste 2 3 4 5 8 7 6 Oasis Waste 2 3 4 5 8 7 6 Oasis Gradient HPLC Narrow Bore Analytical Column (Optional) Mass Spectrometer Alliance HT Mass Spectrometer Analytical Column (optional) Alliance HT Mass Spectrometer Analytical Column (optional) Up to 360 injections were achieved on a single Oasis HLB 2.1mm x 20 mm cartridge column without any visible deterioration of the peak shape. Centrifugation, dilution and/or acidification of the sample prior to analysis may actually provide even longer column lifetimes. Oasis On-Line Column Lifetime 2.1 x 20 mm cartridge column 50 µl injection 0.0 Minutes 1.0 Overlay of Toluamide after 360 Plasma injections. 32

Oasis Prospekt Cartridges On-Line SPE with Oasis Sorbent in Prospekt Cartridges Waters Oasis sorbents are available in Prospekt 1 and Prospekt 2 cartridges for use with the Spark Holland Prospekt /Symbiosis systems. The narrow-bore PVDF cartridge is 1.0 mm x 10 mm contain approximately 2.5 mg of Oasis sorbent. The Oasis sorbents in Prospekt cartridges show the same performance advantage for pharmaceutical compounds over silica-based sorbents. Recovery Comparison of Oasis HLB vs. Bonded Silica Prospekt Cartridge Oasis HLB C 18 C 2 Clonidine % Recovery Terconazole * Data were provided by Dr. Francis Beaudry, a Principal Research Scientist of Phoenix International Life Sciences Inc. in Montreal, Quebec, Canada. SAMPLE PREPARATION Oasis Glass Cartridges for PPT Detection Levels Waters Oasis glass cartridges are available in 5 cc (200 mg) configuration with Teflon frits for trace analysis at parts per trillion level. Each lot is tested for the presence of bisphenol A and other phenols and phthalates, assuring that endocrine disruptors in water samples can be analyzed to part per trillion levels. See the full application in the Oasis Applications Notebook (Waters Literature Code #720000609EN). Endocrine Disruptors PPT Recovery of Estrogens from River Water, LC/MS, 5 ng/l Spike Level, n=4 Results Ions Monitored (m/z) Recovery % RSD 1. bisphenol A 227 113 11 2. 17b-estradiol 271 93 15 3. 17a-ethynylestradiol 295 96 12 4. estrone 269 87 5 5. diethylstilbestrol 267 75 5 Recovery of Phthalates and Nonylphenol from River Water, GC/MS, 200 ng/l Spike Level, n=4 Results Recovery % RSD 1. dimethylphthalate 130 15 2. dirthylphthalate 86 12 3. n-nonylphenol 90 11 4. dibutylphthalate 110 11 5. benzylbutylphthalate 110 8 6. bis(ethylhexyl)phthalate 60 8 ISTD. o-terphenyl (internal standard) 33

Oasis Cartridge Selection Guide 1 cc/ 10 mg 1 cc/ 30 mg 1 cc/30 mg Flangeless 1 cc/30 mg Gilson Adapter 3 cc/ 60 mg 3 cc/60 mg Flangeless 3 cc/60 mg Gilson Adapter 6 cc/ 150 mg NEW NEW Sorbent Box of 100 Box of 100 Box of 100 Box of 500 Box of 100 Box of 100 Box of 500 Box of 30 Oasis HLB 30 µm 186000383 WAT094225 186001879 WAT058882 WAT094226 186001880 WAT058883 N/A Oasis HLB 60 µm N/A N/A N/A N/A N/A N/A N/A N/A Oasis MCX 30 µm N/A 186000252 186001881 186001888 186000254 186001882 N/A 186000256 Oasis MCX 60 µm N/A 186000782 N/A N/A 186000253 N/A N/A 186000255 Oasis MAX 30 µm N/A 186000366 186001883 N/A 186000367 186001884 N/A 186000369 Oasis MAX 60 µm N/A N/A N/A N/A 186000368 N/A N/A 186000370 Oasis 96-well Plates Description 5 mg/ 10 mg/ 30 mg/ 60 mg/ 96-well 96-well 96-well 96-well 1/Pkg 1/Pkg 1/Pkg 1/Pkg Oasis HLB 30 µm 186000309 186000128 WAT058951 N/A Oasis HLB 60 µm N/A N/A N/A 186000679 Oasis MCX 30 µm N/A 186000259 186000248 N/A Oasis MCX 60 µm N/A N/A 186000250 186000678 Oasis MAX 30 µm N/A 186000375 186000373 186001256 Oasis MAX 60 µm N/A N/A N/A 186001205 34

6 cc/ 200 mg 6 cc/400 mg Flangeless 6 cc/ 500 mg 12 cc/ 500 mg 20 cc/ 1 g 35 cc/ 6 g Plus 225 mg Vac RC 30 mg Vac RC 60 mg Glass Cartridge 5 cc/200 mg NEW Box of 30 Box of 100*/500** Box of 30 Box of 20 Box of 20 Box of 10 Box of 50 Box of 50 Box of 50 Box of 30 WAT106202 N/A N/A N/A N/A N/A N/A 186000382 186000381 N/A N/A N/A 186000115 186000116 186000117 186000118 186000132 N/A N/A 186000683 N/A 186001216** N/A N/A N/A N/A N/A N/A 186000261 N/A N/A N/A 186000776 N/A 186000777 186000778 N/A N/A 186000380 N/A N/A 186001855* N/A N/A N/A N/A N/A 186000372 186000371 N/A N/A N/A 186000865 N/A N/A N/A N/A N/A 186000378 N/A SAMPLE PREPARATION Oasis 96-well µelution Plates Waters Manifold for Oasis 96-well Plates Oasis 96-well µelution Plate (Requires Manifold Spacer) Oasis 96-well Plate (No Spacer Required) Description NEW µelution 96-well 1/pkg Oasis HLB 30 µm 186001828BA Oasis MCX 30 µm 186001830BA Oasis MAX 30 µm 186001829 Spacer for µelution Plate (Included with Manifold Kit) Collection Plate Extraction Plate Manifold 186001831 35

Oasis HLB Sample Extraction Products Description Particle Size Quantity Part No. Oasis HLB cartridge 1 cc/10 mg 30 µm 100/box 186000383 Oasis HLB cartridge 1 cc/30 mg 30 µm 100/box WAT094225 NEW Oasis HLB Flangeless cartridge 1 cc/30 mg 30 µm 100/box 186001879 Oasis HLB cartridge with Gilson ASPC Adapter 1 cc/10 mg 30 µm 500/box 186000988 Oasis HLB cartridge with Gilson ASPC Adapter 1 cc/30 mg 30 µm 500/box WAT058882 Oasis HLB cartridge 3 cc/60 mg 30 µm 100/box WAT094226 Oasis HLB cartridge 3 cc/60 mg 30 µm 500/box 186002002 NEW Oasis HLB Flangeless cartridge 3 cc/60 mg 30 µm 100/box 186001880 Oasis HLB cartridge with Gilson ASPC Adapter 3 cc/60 mg 30 µm 500/box WAT058883 Oasis HLB cartridge 6 cc/200 mg 30 µm 30/box WAT106202 Oasis HLB cartridge 6 cc/500 mg 60 µm 30/box 186000115 Oasis HLB cartridge 12 cc/500 mg 60 µm 20/box 186000116 Oasis HLB cartridge 20 cc/1 g 60 µm 20/box 186000117 Oasis HLB cartridge 35 cc/6 g 60 µm 10/box 186000118 Oasis HLB Plus cartridge 225 mg 60 µm 50/box 186000132 Oasis HLB Vac RC cartridge 20 cc/30 mg 30 µm 50/box 186000382 Oasis HLB Vac RC cartridge 20 cc/60 mg 30 µm 50/box 186000381 Oasis HLB Glass cartridge 5 cc/200 mg 60 µm 30/box 186000683 Oasis HLB Prospekt 1 cartridge* 2 mm x 10 mm/15 mg 30 µm 100/box 186000258 NEW Oasis HLB Prospekt 2 cartridge** 1.0 mm x 10 mm 30 µm 96/box 186001196 Reservoir 30 cc for Oasis cartridges 48/box WAT011390 Reservoir 60 cc for Oasis cartridges 12/box WAT024659 Reservoir Adapter for 1 cc, 3 cc, 6 cc cartridges 10/box WAT054260 Reservoir Adapter for 12 cc, 20 cc, 35 cc cartridges 10/box WAT048160 Reservoir Adapter for 5 cc cartridges, Teflon 10/pkg 405000934 NEW Oasis HLB column 2.1 mm x 20 mm 5 µm 1/pkg 186002034 NEW Oasis HLB column 3.0 mm x 20 mm 5 µm 1/pkg 186002037 NEW Oasis HLB column 3.9 mm x 20 mm 5 µm 1/pkg 186002040 Oasis HLB cartridge column 3.9 mm x 20 mm 5 µm 1/pkg 186001413 NEW Oasis HLB 4.6 mm x 20 mm 5 µm 1/pkg 186002043 NEW Oasis HLB column 2.1 mm x 20 mm 15 µm 1/pkg 186002035 NEW Oasis HLB column 3.0 mm x 20 mm 15 µm 1/pkg 186002038 NEW Oasis HLB column 3.9 mm x 20 mm 15 µm 1/pkg 186002041 Oasis HLB cartridge column 3.9 mm x 20 mm 15 µm 1/pkg 186001414 NEW Oasis HLB column 4.6 mm x 20 mm 15 µm 1/pkg 186002044 Oasis HLB column 2.0 mm x 15 mm 25 µm 1/pkg 186001792 NEW Oasis HLB column 2.1 mm x 20 mm 25 µm 1/pkg 186002036 Oasis HLB cartridge column 2.1 mm x 20 mm 25 µm 1/pkg 186000706 NEW Oasis HLB column 3.0 mm x 20 mm 25 µm 1/pkg 186002039 NEW Oasis HLB column 3.9 mm x 20 mm 25 µm 1/pkg 186002042 NEW Oasis HLB column 4.6 mm x 20 mm 25 µm 1/pkg 186002045 Holder Kit for 2.1 mm x 20 mm cartridge column 1/pkg 186000262 Holder Kit for 3.9 mm x 20 mm cartridge column 1/pkg WAT046910 Extraction Column Connector 1/pkg WAT082745 Inline precolumn filter kit 1/pkg WAT084560 Replacement filters 5/pkg WAT005139 Replacement steel gaskets 1/pkg WAT084567 Oasis HLB µelution Plate 96-well 30 µm 1/pkg 186001828BA Oasis filter plate 96-well 24/pkg 186001136 Oasis HLB plate 5 mg/96-well 30 µm 1/pkg 186000309 Oasis HLB plate 10 mg/96-well 30 µm 1/pkg 186000128 Oasis HLB plate 30 mg/96-well 30 µm 1/pkg WAT058951 Oasis HLB plate 60 mg/96-well 60 µm 1/pkg 186000679 * For use with Spark Holland Prospekt systems ** For use with Spark Holland Symbiosis system 36

Oasis MCX Sample Extraction Products (Cation-Exchanger) Description Particle Size Quantity Part No. Oasis MCX cartridge 1 cc/30 mg 30 µm 100/box 186000252 NEW Oasis MCX Flangeless cartridge 1 cc/30 mg 30 µm 100/box 186001881 Oasis MCX cartridge 1 cc/60 mg 60 µm 100/box 186000782 Oasis MCX cartridge 3 cc/60 mg 30 µm 100/box 186000254 NEW Oasis MCX Flangeless cartridge 3 cc/60 mg 30 µm 100/box 186001882 Oasis MCX cartridge 3 cc/60 mg 60 µm 100/box 186000253 Oasis MCX cartridge 6 cc/150 mg 30 µm 30/box 186000256 Oasis MCX cartridge 6 cc/150 mg 60 µm 30/box 186000255 Oasis MCX cartridge 6 cc/500 mg 60 µm 30/box 186000776 Oasis MCX cartridge 20 cc/1 g 60 µm 20/box 186000777 Oasis MCX cartridge 35 cc/6 g 60 µm 10/box 186000778 Oasis MCX Vac RC cartridge 20 cc/60 mg 30 µm 50/box 186000261 Oasis MCX Vac RC cartridge 20 cc/60 mg 60 µm 50/box 186000380 NEW Oasis MCX Prospekt 2 cartridge** 10 mm x 1 mm 30 µm 96/box 186002098 NEW Oasis MCX direct connect column 2.1 mm x 15 mm 30 µm 1/pkg 186002050 NEW Oasis MCX column 2.1 mm x 20 mm 30 µm 1/pkg 186002046 NEW Oasis MCX cartridge column 2.1 mm x 20 mm 30 µm 1/pkg 186002051 NEW Oasis MCX column 3.0 mm x 20 mm 30 µm 1/pkg 186002047 NEW Oasis MCX column 3.9 mm x 20 mm 30 µm 1/pkg 186002048 NEW Oasis MCX column 4.6 mm x 20 mm 30 µm 1/pkg 186002049 Oasis MCX µelution Plate 96-well 30 µm 1/pkg 186001830BA Oasis MCX plate 10 mg/96-well 30 µm 1/pkg 186000259 Oasis MCX plate 30 mg/96-well 30 µm 1/pkg 186000248 Oasis MCX plate 30 mg/96-well 60 µm 1/pkg 186000250 Oasis MCX plate 60 mg/96-well 60 µm 1/pkg 186000678 SAMPLE PREPARATION Oasis MAX Sample Extraction Products (Anion-Exchanger) Description Particle Size Quantity Part No. Oasis MAX cartridge 1 cc/30 mg 30 µm 100/box 186000366 NEW Oasis MAX Flangeless cartridge 1 cc/30 mg 30 µm 100/box 186001883 Oasis MAX cartridge 3 cc/60 mg 30 µm 100/box 186000367 Oasis MAX cartridge 3 cc/60 mg 60 µm 100/box 186000368 NEW Oasis MCX Flangeless cartridge 3 cc/60 mg 60 µm 100/box 186001884 Oasis MAX cartridge 6 cc/150 mg 30 µm 30/box 186000369 Oasis MAX cartridge 6 cc/150 mg 60 µm 30/box 186000370 Oasis MAX cartridge 6 cc/500 mg 60 µm 30/box 186000865 Oasis MAX Vac RC cartridge 20 cc/30 mg 30 µm 50/box 186000372 Oasis MAX Vac RC cartridge 20 cc/60 mg 30 µm 50/box 186000371 Oasis MAX Vac RC cartridge 20 cc/60 mg 60 µm 50/box 186000378 NEW Oasis MAX Prospekt 2 cartridge** 10 mm x 1 mm 30 µm 96/box 186002099 NEW Oasis MAX direct connect column 2.1 mm x 15 mm 30 µm 1/pkg 186002056 NEW Oasis MAX column 2.1 mm x 20 mm 30 µm 1/pkg 186002052 NEW Oasis MAX cartridge column 2.1 mm x 20 mm 30 µm 1/pkg 186002057 NEW Oasis MAX column 3.0 mm x 20 mm 30 µm 1/pkg 186002053 NEW Oasis MAX column 3.9 mm x 20 mm 30 µm 1/pkg 186002054 NEW Oasis MAX column 4.6 mm x 20 mm 30 µm 1/pkg 186002055 Oasis MAX µelution Plate 96-well 1/pkg 186001829 Oasis MAX plate 10 mg/96-well 30 µm 1/pkg 186000375 Oasis MAX plate 30 mg/96-well 30 µm 1/pkg 186000373 Oasis MAX plate 60 mg/96-well 30 µm 1/pkg 186001256 Oasis MAX plate 60 mg/96-well 60 µm 1/pkg 186001205 ** For use with Spark Holland Symbiosis system 37

Oasis Method Development Kit Description Particle Size Quantity Part No. Oasis Method Development Kit 3 cc/60 mg 30 µm 10 each MCX, HLB, MAX 186000867 Manifold for Extraction Plate Description Quantity Part No. Extraction plate manifold for Oasis 96-well plates 1/box 186001831 Extraction Plate Manifold Kit A WAT097944 (includes extraction plate manifold, reservoir tray, sealing cap and 350 µl sample collection plate) Extraction Plate Manifold Kit B WAT097945 (as kit A, with 1 ml sample collection plate) Extraction Plate Manifold Kit C WAT097946 (as kit A, with 2 ml sample collection plate) Accessories for Extraction Plate Manifold Description Quantity Part No. Disposable Reservoir tray 25/box WAT058942 Sample collection plate, 350 µl 50/box WAT058943 Sample collection plate, 2 ml 50/box WAT058958 Sealing cap for 96-well collection plate 50 sheets/pkg WAT058959 SPE Vacuum Pump 115V 60 Hz 725000417 SPE Vacuum Pump 240V 50 Hz 725000418 Oasis 96-well µelution Plate (Requires Manifold Spacer) Spacer for µelution Plate (Included with Manifold Kit) Collection Plate Oasis 96-well Plate (No Spacer Required) Extraction Plate Manifold 186001831 Manifold for Extraction Cartridges Description Waters extraction manifold, 20-position without rack (includes 20 needle tips, 25 plugs, and ejector tool) Waters extraction manifold, 20-position (complete with rack for 13 mm x 75 mm tubes) Waters extraction manifold, 20-position (complete with rack for 13 mm x 100 mm tubes) Waters extraction manifold, 20-position (complete with rack for 16 mm x 75 mm tubes) Waters extraction manifold, 20-position (complete with rack for 16 mm x 100 mm tubes) Part No. WAT200677 WAT200606 WAT200607 WAT200608 WAT200609 Accessories for Extraction Columns and Cartridges Description Quantity Part No. Holder Kit for 2.1 mm x 20 mm cartridge column 1/pkg 186000262 Holder Kit for 3.9 mm x 20 mm cartridge column 1/pkg WAT046910 Extraction Column Connector 1/pkg WAT082745 Inline precolumn filter kit 1/pkg WAT084560 Replacement filters 5/pkg WAT005139 Replacement steel gaskets 1/pkg WAT084567 SPE Vacuum Pump 115V 60 Hz 725000417 SPE Vacuum Pump 240V 50 Hz 725000418 38

NEW Sep-Pak 96-Well Plates The 96-well plate has two stage diameter well design. This design allows for varying quantities of sorbent within the well. The lower portion of the well has a smaller diameter which accommodates smaller sorbent masses, packed to desirable aspect ratios. This means the sorbent is packed to desirable height to width ratios, resulting in good sample distribution and bed depth. The upper portion of the well has a larger diameter. Both well diameters are used for larger sorbent masses, resulting in good sample distribution, bed height and sample load for larger sample sizes. Sep-Pak 96-Well Plates are now available with chemistries that have been proven for years in Sep-Pak Cartridges. These plates have been introduced for high volume applications where the need to process 100 s of samples is a requirement. Plates can be used on most vacuum manifolds, robotic and automated sample handling systems. Sep-Pak Plates are available with different sorbent masses for different sample loads. Sep-Pak 96-Well Plates are extensively tested for quality control. Each plate is shipped with a certificate of analysis that covers tests performed on raw silica and the bonded silica phase. Each lot is tested for bonding, cleanliness and chromatographic selectively. The plates are packed and tested to ensure consistent flow properties and sorbent mass across the 96 wells. Waters 96-Well Plate Design By varying frit size and/or placement, the same plate may be filled with various quantities of sorbent per well. Our design permits optimal recoveries, even with low sorbent weights for smaller elution volumes. SAMPLE PREPARATION 5 mg 10 mg 60 mg Sep-Pak tc 18 is now available in the patented* µelution Plate. This plate is designed for small volume samples. The recommended protocol for elution from this plate is as low as 25 µl. The low volume extracts from the plate can be injected directly, eliminating time consuming evaporation steps. Sep-Pak 96-Well Plates Description Part No. Sep-Pak tc 18 µelution Plate 186002318 Sep-Pak tc 18 25 mg Plate 186002319 Sep-Pak tc 18 40 mg Plate 186002320 Sep-Pak tc 18 100 mg Plate 186002321 Sep-Pak Accell Plus QMA 100 mg Plate 186001917 * Patent Pending 39

Solid-Phase Extraction Read the Primer on SPE - available online. Which Cartridge Design is Best for Your Application Sep-Pak cartridges are available in a variety of designs; each offers specific functional benefits but all contain the same high-quality packing sorbents. This makes transfer of methods from one design to another straightforward and predictable. Sep-Pak Plus Cartridges for Maximum Versatility Sep-Pak Plus cartridges are designed for both convenient manual use and compatibility with automated instruments. They come in short body and long body configurations and are constructed from high purity polyethylene with female Luer inlets and male Luer outlets. They can be connected to many different types of devices such as positive flow pumps and syringes, vacuum manifolds, and automated sample processors. In many applications, the use of positive flow (pumps) provides for increased reproducibility in your results. Variations in recovery data, due to flow rate variability and the potential for air drying out a cartridge when using vacuum devices, can be eliminated. The male outlet tips feature reduced hold-up volume for precise fraction collection. The fittings also allow cartridges to be easily stacked for increased capacity or for the use of more than one sorbent type for cleanup of complex sample mixtures. Sep-Pak Plus cartridges can be used with liquid or gas samples and have the unique capability for reverse flow direction, a useful feature for trace enrichment applications. The packing process and compression seal provide a uniform bed structure to minimize band spreading for the best possible separation efficiency. Bed volumes and weights of sorbent in the short and long body cartridges equal those of the corresponding Classic cartridges. All Sep-Pak Plus cartridges have color-coded compression rings for general sorbent identification. Sep-Pak Classic Cartridges Sep-Pak Classic cartridges were the original SPE cartridges introduced 25 years ago. Waters Sep-Pak Classic cartridges contain the same amount of sorbent as the Sep-Pak Plus cartridge. This means that you can convert existing methods easily to the more convenient Plus configuration. Both the Sep-Pak Plus and Classic designs have two different sizes (short body and long body) depending on the type of sorbent. Sep-Pak Light Cartridges for Limited Sample Volumes/Lower Solvent Consumption Sep-Pak Light cartridges look similar to the short-body Sep-Pak Plus cartridges, except for the distinctive finned outer body. The functional difference is the reduced internal diameter which results in bed volumes about one-third that of the corresponding Plus cartridge. This design allows you to elute fractions in a minimal volume to improve recoveries and reduce solvent consumption, a benefit for sample-limited applications or where excessive dilution is a concern. In addition, the Sep-Pak Light cartridges are specially designed to provide efficient packed sorbent beds for use in downsizing current applications to reduce solvent consumption. All Sep-Pak Light cartridges have color-coded compression rings for general sorbent identification. Sep-Pak Vac RC Cartridges for Manual or Robotic Sample Preparation These cartridges feature a 20 cc funnel shape design containing 100 or 500 mg of sorbent. They can be used with robotic sample preparation equipment or with vacuum manifolds. The integral reservoir design allows for the convenient processing of larger amounts of sample or solvents. This reduces setup time and costs associated with the use of separate reservoirs. Color-coded printing on the cartridges makes sorbent identification easy. Economical Sep-Pak Vac Cartridges Sep-Pak Vac cartridges provide you with the option of even lower cost sample preparation. The cartridges use molded polypropylene syringebarrel bodies and two polyethylene filters to contain the packing material. A standard male Luer outlet tip permits use with most multiposition vacuum manifolds. Cartridge sizes range from 1 cc to 35 cc and sorbent weights range from 50 mg up to 10 grams. Sizes up to 6 cc can be used on many automated sample preparation systems designed to use syringe-barrel style cartridges. Sep-Pak Vac cartridges offer the highest quality at low cost with a complete range of capacities for use in optimizing your methods. Color-coded printing on the syringe-barrel cartridges makes sorbent identification easy. Sep-Pak C 18 Environmental Cartridges This is a special long body Plus cartridge design containing approximately 1 gram of the monofunctional C 18 or the trifunctional C 18 sorbent. These cartridges are especially useful in trace enrichment applications. 40

The Anatomy of Sep-Pak Plus and Classic Cartridge Design The Anatomy of Sep-Pak Vac and Vac RC Cartridge Designs Sep-Pak Plus Sep-Pak Classic Sep-Pak Vac Sep-Pak Vac RC 1 7 2 3 4 5 5 1. Female Luer inlet accepts male Luer tip. Plus cartridge design can be stacked. 2. Highest quality sorbents designed and made specifically for sample preparation; clean, dry, reproducible in activity and capacity with optimal surface area, pore, and particle size distributions. 3. Broad range of sorbent surface activities available; each lot is tested under rigid specifications for chromatographic activity, retention and selectivity. 4. Sorbent type and bed dimensions equal to corresponding Sep-Pak Classic cartridges enable direct transfer of previously developed and published methods to new Plus design. 5. Weight of sorbent in each cartridge is controlled within +/- 5% of specification to assure reproducible performance. 6 6 7 11 10 8 9 6*. Advanced bed formation technology to minimize voids and channels. Patented Radial Compression Technology used to form homogeneous packed bed free of voids and channels. * 1. Highest quality sorbents designed and made specially for sample preparation; clean, dry, reproducible in activity and capacity, with optimal surface area pore and particle size distributions. 2. Broad range of sorbent surface activities available; each lot is tested under rigid specifications for chromatographic activity, retention and selectivity. 3. Weight of sorbent in each cartridge is controlled within +/- 5% of specification to assure reproducible performance. 4. Special blend of HD and UHMW polyethylenes used for 20 micron frits. 5. Molded, medical grade polypropylene body. 6. Cartridges are sealed in a special polyfoil pouch to protect product integrity, sorbent activity and purity. 7. Integral reservoir approximately 20 ml, robotic compatible. 8. Outlet male Luer tip. 1 2 3 4 8 6 9 SAMPLE PREPARATION 7. Special blend of HD and UHMW polyethylenes used for 20 micron frits imparts excellent solvent resistance, extremely low extractables level, and good flow properties. Frit also acts as depth filter for small amounts of sample debris. 8. Polyethylene body has excellent solvent resistance. All body parts are quality tested to verify extremely low level of UV-absorbing extractables. Plus design is molded for precise dimensions making it suitable for automated equipment. 9. Cartridges are sealed in a special polyfoil pouch to protect product integrity, sorbent activity and purity. 10. Male Luer outlet has reduced internal volume for minimal sample hold-up. 11. Color-coded ring compresses and seals the cartridge and identifies sorbent. 9. Color-coded labeling on the cartridge to identify the sorbent. * P.D. McDonald, C. W. Rausch, Radial Compression of Packed Beds, U.S. Patent # 4,250,035 (1981); Great Britain # 1,569,700 (1976); Canada # 1,101,785 (1981); Japan # 1,400,983 (1987); Sweden # 450,750 & # 453,437 (1987); Germany # 2,655,650 (1988); other patents pending. 41

Sep-Pak Cartridge Selection Guide Plus Classic Light Vac 1 cc 50 mg Vac 1 cc 100 mg Vac RC 100 mg Vac 3 cc 200 mg Waters Sep-Pak Vac Waters Sep-Pak Vac Waters Sep-Pak Vac Sorbent Box of 50 Box of 50 Box of 50 Box of 100 Box of 100 Box of 50 Box of 50 C 18 WAT020515 WAT051910 WAT023501 WAT054955 WAT023590 WAT036935 WAT054945 tc 18 WAT036810 N/A WAT036805 WAT054960 WAT036820 WAT043410 WAT054925 C 8 WAT036775 N/A WAT036770 WAT054965 WAT036785 WAT043415 WAT054940 tc 2 WAT052720 N/A WAT052725 N/A WAT052710 N/A N/A Silica WAT020520 WAT051900 WAT023537 WAT054980 WAT023595 WAT036940 WAT054930 Florisil WAT020525 WAT051960 WAT023543 WAT054985 WAT023600 N/A N/A Accell Plus CM WAT020550 WAT010910 WAT023531 N/A WAT023625 N/A N/A Accell Plus QMA WAT020545 WAT010835 WAT023525 N/A WAT023620 WAT043460 N/A Alumina A WAT020500 WAT051800 WAT023549 N/A WAT023575 N/A N/A Alumina B WAT020505 WAT051820 WAT023555 N/A WAT023580 N/A N/A Alumina N WAT020510 WAT051810 WAT023561 N/A WAT023585 N/A N/A Amino Propyl (NH 2 ) WAT020535 WAT010830 WAT023513 N/A WAT023610 WAT043475 N/A Cyano Propyl (CN) WAT020540 WAT010823 WAT023507 WAT054975 WAT023615 WAT043470 WAT054935 Diol WAT020530 WAT010818 WAT023519 N/A WAT023605 WAT043480 N/A NEW Sep-Pak 96-well Plates Description NEW Sep-Pak µelution 96-well 1/pkg Sep-Pak tc 18 25 mg Plate 186002319 Sep-Pak tc 18 40 mg Plate 186002320 Sep-Pak tc 18 100 mg Plate 186002321 Sep-Pak Accell Plus QMA 100 mg Plate 186001917 42

Vac 3 cc 500 mg Vac RC 500 mg Vac 6 cc 500 mg Vac 6 cc 1 gram Vac 12 cc 2 gram Vac 20 cc 5 gram Vac 35 cc 10 gram Waters Sep-Pak Vac Waters Sep-Pak Vac Box of 50 Box of 50 Box of 30 Box of 30 Box of 20 Box of 20 Box of 10 Waters Sep-Pak Vac WAT020805 WAT036945 WAT043395 WAT036905 WAT036915 WAT036925 WAT043345 C 18 WAT036815 WAT043425 WAT036790 WAT036795 WAT043380 WAT043365 WAT043350 tc 18 WAT036780 WAT043430 WAT054525 WAT054570 WAT054615 WAT054660 WAT054700 C 8 Waters Sep-Pak Vac Waters Sep-Pak Vac Waters Sep-Pak Vac Sorbent SAMPLE PREPARATION WAT052715 N/A N/A WAT052705 N/A N/A N/A tc 2 WAT020810 WAT036950 WAT043400 WAT036910 WAT036920 WAT036930 WAT043355 Silica WAT020815 WAT043435 WAT043405 WAT043390 WAT043385 WAT043370 WAT043360 Florisil WAT020855 WAT054505 WAT054545 WAT054590 WAT054635 WAT054675 WAT054720 Accell Plus CM WAT020850 WAT054500 WAT054550 WAT054595 WAT054640 WAT054680 WAT054725 Accell Plus QMA WAT020820 N/A WAT054535 WAT054580 WAT054620 WAT054670 WAT054710 Alumina A WAT020825 N/A WAT054540 WAT054585 WAT054625 WAT054665 WAT054715 Alumina B WAT020830 WAT043485 WAT054530 WAT054575 WAT054630 WAT043375 WAT054705 Alumina N WAT020840 WAT054515 WAT054560 WAT054605 WAT054650 WAT054695 WAT054740 Amino Propyl (NH 2 ) WAT020835 N/A WAT054555 WAT054600 WAT054645 WAT054685 WAT054730 Cyano Propyl (CN) WAT020845 WAT054520 WAT054565 WAT054610 WAT054655 WAT054690 WAT054735 Diol NEW Sep-Pak 96-well µelution Plate Description NEW Sep-Pak µelution 96-well 1/pkg Sep-Pak tc 18 µelution Plate 186002318 mg = nominal weight of sorbent in milligrams. See Sep-Pak cartridge configuration and nominal weight of sorbent chart on page 46 for specific values of each sorbent. 43

Which Sorbent is Best For Your Application Selecting the most appropriate sorbent is best approached from both a theoretical and empirical standpoint. Knowledge of the components in a specific sample will help you determine the general mode of separation, such as, normal phase, reversed-phase, or ion exchange, and more specific knowledge may help you select between the various types of sorbents in each mode. However, it is frequently necessary to determine the optimal separation by empirical means. The text and tables that follow give you detailed information on the properties of the sorbents that Waters offers. The Original C 18 Sorbent Waters original C 18 sorbent is the most widely referenced SPE product, with over 2,500 published papers or articles specifying its use. This product is made using the most reproducible bonding technology available, utilizing monofunctional silane chemistry. Waters original C 18 sorbent offers a unique selectivity and excellent recovery of analytes, and is recommended for most applications where HPLC is utilized for subsequent analysis. tc 18 (trifunctional) Sorbent With the addition of tc 18 sorbent, you can now choose between two highly reproducible C 18 chemistries, monofunctional and trifunctional. Trifunctional tc 18 cartridges are recommended for most applications involving GC, GC/MS or LC/MS especially if long sample contact times are involved. This new tc 18 sorbent has also become increasingly popular in pharmaceutical applications when a high carbon load, very hydrophobic surface is required. (See chart). tc 2 (trifunctional) Sorbent Many applications require a less hydrophobic (relative to C 18 ), short carbon chain sorbent to provide optimal sample processing of a wide range of analytes. However, finding a reproducible, stable C 2 sorbent can be difficult. Waters has recently developed a new, state-of-the-art trifunctional bonding process that provides you with the most dependable sorbents (tc 2 and tc 18 ) of this type available. The tc 2 is ideal for many types of applications such as environmental and pharmaceutical. Recommendations for Using C 18 and tc 18 Sorbents C 18 tc 18 Sample Contact Time long (> 10 minutes) NR R short (< 10 minutes) R O Sample ph < 3 NR R 3-7 R O > 7 NR R Detection Method HPLC Ultraviolet/Visible Absorbance R O Fluorescence R O GC Flame Ionization O R Mass Selective O R Electron Capture O R Nitrogen-Phosphorous O R Selectivity Unique R NR Similar to other vendors C 18 cartridges NR R Key: R = recommended sorbent O = may also be applicable NR = not recommended Sep-Pak Cartridges: Chemistry of Bonded-Phase Sorbents Sorbent Chemistry Bonding Chemistry Surface Functionality C 18 Monofunctional silane Si(CH 3 ) 2 C 18 H 37 tc 18 Trifunctional silane SiC 18 H 37 C 8 Monofunctional silane Si(CH 3 ) 2 C 8 H 17 tc 2 Trifunctional silane -SiC 2 H 5 NH 2 Trifunctional silane Si(CH 2 ) 3 NH 2 CN Difunctional silane Si(CH 3 )(CH 2 ) 3 CN Diol Trifunctional silane Si(CH 2 ) 3 OCH 2 CH(OH)CH 2 OH Accell Plus CM Polymeric* CO 2 - Na+ Accell Plus QMA Polymeric** C(O)NH(CH 2 ) 3 N(CH 3 ) 3 + Cl- DNPH-Silica Coated*** * An acrylic acid/acrylamide copolymer on Diol silica; ligand density: 350 µmoles/g; approximate protein binding capacity = 175 mg Cytochrome c/g of sorbent or 63 mg/accell Plus CM Classic or Plus cartridge. ** An acrylamide copolymer on Diol silica; ligand density: 220 µmoles/g; approximate protein binding capacity = 200 mg BSA/g of sorbent or 72 mg/accell Plus QMA Classic or Plus cartridge. *** Unbonded silica sorbent coated with 14 µmoles DNPH/g of sorbent or 5 µmoles/cartridge. 44

Sorbent Properties and Typical Applications Separation Mode Sorbent Reversed-Phase C 18 Properties & Applications silica-based bonded phase with strong hydrophobicity; used to adsorb analytes of even weak hydrophobicity from aqueous solutions; typical applications include drugs and their metabolites in serum, plasma or urine, desalting of peptides, trace organics in environmental water samples, organic acids in beverages; similar in behavior to reversed-phase HPLC columns tc 18 silica-based bonded phase with strong hydrophobicity; trifunctional bonding chemistry gives it an increased hydrolytic stability over C 18 ; applications similar to C 18 C 8 silica-based bonded phase with moderate hydrophobicity; use for methods requiring less retention than C 18 ; typical applications include drugs and their metabolites in serum, plasma or urine, peptides in serum, plasma tc 2 silica-based bonded phase with low hydrophobic character; use for methods requiring less retention than C 8 ; applications are similar to C 18 and C 8 Porapak RDX specially cleaned polymeric poly(divinylbenzene-vinylpyrrolidone) resin with hydrophobic character. Can be used as an alternative to octadecyl-bonded silica for preparation of analytes that weakly adsorb to silica-based reversed-phase sorbents. Compatible with sample or eluents at high and low ph. Specifically designed for the concentration of high explosives in aqueous samples. Reversed or Normal Phase: Amino Propyl (NH 2 ) Cyano Propyl (CN) Diol Normal Phase Silica Alumina (A, B & N) Florisil Ion-Exchange Accell Plus QMA Accell Plus CM Specialty DNPH-Silica, XPoSure silica-based polar bonded phase with basic character; can be used as a polar sorbent, like silica, with different selectivity for acidic/basic analytes or as weak anion exchanger in aqueous medium; applications include phenols and phenolic pigments, petroleum fractionation, saccharides and drugs and metabolites silica-based polar bonded phase; can be used as less polar alternative to silica in normal-phase applications or as less hydrophobic alternative to C 18 or C 8 in reversed-phase applications; typical applications include drugs, drug metabolites and pesticides silica-based polar bonded phase with neutral character; can be used as an alternative to silica in normal phase applications, where the acidic character of silica is undesirable or as very weakly interacting phase in aqueous applications; applications include antibiotics from cosmetics; isolation of proteins or peptides by hydrophobic interaction chromatography polar sorbent, used primarily to adsorb analytes from non polar solvents like hydrocarbons, chloro- or fluoro-substituted hydrocarbons or less polar esters and ethers; elution with more polar solvents like polar esters, ethers, alcohols, acetonitrile or water; the binding mechanism can be hydrogen bonding or dipole-dipole interaction; silica can also be used in aqueous medium as a cation exchanger of intermediate strength. similar in use to silica; available in acidic, basic and neutral high activity grades; alumina also exhibits specific interactions with the pi-electrons of aromatic hydrocarbons, making it useful for applications like crude oil fractionation; acidic and basic grades can also be used as low-capacity ion-exchangers polar, highly active, weakly basic sorbent for adsorption of low to moderate polarity species from nonaqueous solutions; specifically designed for the adsorption of pesticides using official AOAC and EPA methods; other applications include polychlorinated biphenyls (PCB s) in transformer oil silica-based hydrophilic strong anion-exchanger with large pore-size; extraction of anionic analytes in aqueous and non-aqueous solutions; due to the large pore-size, it is excellent for the isolation of anionic proteins, e.g., immunoglobulins, enzymes; other applications include the removal of acidic pigments from wines, fruit juices and food extracts, isolation of phenolic compounds, peptide pool fractionations silica-based hydrophilic weak cation-exchanger with large pore-size; extraction of cationic analytes in aqueous and non-aqueous solutions; due to the large pore-size, it is excellent for the isolation of cationic proteins; other applications include pesticides, herbicides, steroids contains acidified dinitrophenylhydrazine reagent coated on a silica sorbent. Used for the collection of air samples and subsequent quantitation of aldehydes and ketones by reaction to form the hydrazone derivative, and analysis by HPLC. DNPH-Silica is specified in several EPA procedures for the analysis of carbonyl compounds in air. SAMPLE PREPARATION 45

Sep-Pak Cartridge Sorbents Physicochemical Characteristics Nominal Weight of Sorbent (in milligrams) Activity End- Percent Pore Size Particle Size Sorbent Grade capped Carbon (nominal, Å) Range (µm) C 18 N/A Yes 12 125 55-105 tc 18 N/A Yes 17 125 37-55 C 8 N/A Yes 9 125 37-55 tc 2 N/A Yes 2.7 125 37-55 NH 2 N/A No 3.5 125 55-105 CN N/A Yes 6.5 125 55-105 Diol N/A No 2 300 37-55 Silica High* N/A N/A 125 55-105 Florisil High** N/A N/A 60 50-200 Alumina A 1*** N/A N/A 120 50-300 Alumina N 1*** N/A N/A 120 50-300 Alumina B 1*** N/A N/A 120 50-300 Accell Plus CM N/A N/A 5.5 300 37-55 Accell Plus QMA N/A N/A 6 300 37-55 Porapak RDX N/A N/A 95 200 125-150 DNPH-Silica N/A N/A N/A 125 55-105 XPoSure N/A N/A N/A 125 500-1000 * Moisture content 3.2% ** Moisture content 2.5% *** Moisture content 1.5%; activity 1 on Brockmann activity scale Separation Mode Cartridge Type General Surface and Sorbent Plus Classic Light Chemistry Reversed-phase C 18 360 360 130 Neutral Environmental C 18 * 820 tc 18 400 N/A 145 Neutral Environmental tc 18 * 900 C 8 400 N/A 145 Neutral tc 2 400 N/A 145 Neutral Normal or Reversed-phase Amino Propyl (NH 2 )** 360 360 130 Basic Cyano Propyl (CN) 360 360 130 Neutral Diol 360 360 130 Neutral Normal Phase Silica 690 690 120 Neutral Florisil 910 900 145 Basic Alumina A 1710 1850 280 Acidic Alumina B 1710 1850 280 Basic Alumina N 1710 1850 280 Neutral Ion-Exchange ** Accell Plus QMA 360 360 130 Basic Accell Plus CM 360 360 130 Acidic *Sep-Pak Plus long-body cartridge. **The amino propyl packing may also be used in the protonated form as a weak anion exchanger. Sep-Pak Vac Cartridges Nominal Weight of Sorbent Syringe Size 50 mg 100 mg 200 mg 500 mg 1 g 2 g 5 g 10 g 1 cc X X Vac RC X 3 cc X X Vac RC X 6 cc X X 12 cc X 20 cc X 35 cc X See Cartridge Selection Guide for specific product availability. 46

Sep-Pak Cartridges Sep-Pak Plus Sep-Pak Classic Sep-Pak Light Box of 50 Cartridges Cartridges Cartridges C 18 WAT020515 WAT051910 WAT023501 C 18 Environmental Cartridge (long body) WAT023635 tc 18 (trifunctional) WAT036810 WAT036805 tc 18 Environmental Cartridge (long body) WAT036800 C 8 WAT036775 WAT036770 tc 2 (trifunctional) WAT052720 WAT052725 Silica WAT020520 WAT051900 WAT023537 Florisil WAT020525 WAT051960 WAT023543 Accell Plus CM WAT020550 WAT010910 WAT023531 Accell Plus QMA WAT020545 WAT010835 WAT023525 Alumina A WAT020500 WAT051800 WAT023549 Alumina B WAT020505 WAT051820 WAT023555 Alumina N WAT020510 WAT051810 WAT023561 Amino Propyl (NH 2 ) WAT020535 WAT010830 WAT023513 Cyano Propyl (CN) WAT020540 WAT010823 WAT023507 Diol WAT020530 WAT010818 WAT023519 Sep-Pak Plus cartridges come in short body (C 18, tc 18, C 8, tc 2, Accell Plus QMA and CM, NH 2, CN, Diol) and long body (Silica, Florisil, Alumina A,B,N) configurations. They are stackable, allowing for increased capacity or for the use of more than one sorbent. Sep-Pak Plus cartridges are compatible with some automated sample processors. SAMPLE PREPARATION Sep-Pak Plus Methods Development Kits Description Quantity Part No. Methods Development Kit A Box 100 WAT039545 (contains 10 Sep-Pak cartridges each of Alumina: Acid, Basic, Neutral, CN, Diol, Silica, Florisil, Accell Plus QMA, C 18, and NH 2 ) Methods Development Kit B Box 50 WAT039555 (contains 10 Sep-Pak cartridges each of NH 2, CN, Diol, Silica and C 18 ) Methods Development Kit C Box 50 WAT039565 (contains 10 Sep-Pak cartridges each of Accell Plus QMA, Accell Plus CM, C 18, Silica and Diol) Sep-Pak Classic cartridges also come in short body (C 18, Accell Plus QMA and CM, NH 2, CN, Diol) and long body (Silica, Florisil, Alumina A,B,N) configurations. Waters original Sep-Pak cartridges, radially compressed mini-columns, for versatile easy column liquid-solid extraction. Sep-Pak Cartridge Experiment Kit This activity is designed to demonstrate the principles of liquid chromatography using a grape drink mix and Sep-Pak C 18 Plus cartridges. This experiment lends itself ideally to a classroom setting. Description Cartridge Experiment Kit (Includes two Sep-Pak C 18 cartridges and 1 pack of grape mix with instructions) Part No. WAT088253 For sample limited applications or where excessive dilution is a concern, Sep-Pak Light cartridges contain less sorbent than the standard Sep-Pak cartridge and can give higher sample recovery with less solvent consumption for limited sample situations. 47

Sep-Pak Vac SPE Cartridges Waters Sep-Pak Vac solid-phase extraction columns come in a wide range of syringe-barrel configurations, from 1 cc to 35 cc, for all your high volume batch processing and automated sample prep needs. They are packed with a variety of sorbent weights from 50 mg to 10 grams to handle all your sample load requirements. Sep-Pak Vac cartridges provide the method reproducibility you need for dependable analytical results and they are registered with the FDA as Class 1 medical Devices. They are manufactured in strict compliance with cgmp guidelines in manufacturing facilities certified to meet the ISO 9002 standard. Sep-Pak Vac 1 cc Cartridges (contains 50 mg of sorbent) Sep-Pak Vac 3 cc Cartridges (contains 500 mg of sorbent) Waters Sep-Pak Vac Box of 100 C 18 tc 18 (trifunctional) C 8 Silica Florisil Cyano Propyl (CN) Part No. WAT054955 WAT054960 WAT054965 WAT054980 WAT054985 WAT054975 For information on the availability of other sorbents, please contact your local sales representative. Sep-Pak Vac 1 cc Cartridges (contains 100 mg of sorbent) Waters Sep-Pak Vac Box of 100 C 18 tc 18 (trifunctional) C 8 tc 2 (trifunctional) Silica Florisil Accell Plus CM Accell Plus QMA Alumina A Alumina B Alumina N Amino Propyl (NH 2 ) Cyano Propyl (CN) Diol Sep-Pak Vac 3 cc Cartridges (contains 200 mg of sorbent) Waters Sep-Pak Vac Box of 50 C 18 tc 18 (trifunctional) C 8 Silica Cyano Propyl (CN) Part No. WAT023590 WAT036820 WAT036785 WAT052710 WAT023595 WAT023600 WAT023625 WAT023620 WAT023575 WAT023580 WAT023585 WAT023610 WAT023615 WAT023605 Part No. WAT054945 WAT054925 WAT054940 WAT054930 WAT054935 Waters Sep-Pak Vac Box of 50 C 18 tc 18 (trifunctional) C 8 tc 2 (trifunctional) Silica Florisil Accell Plus CM Accell Plus QMA Alumina A Alumina B Alumina N Amino Propyl (NH 2 ) Cyano Propyl (CN) Diol Sep-Pak Vac 6 cc Cartridges (contains 500 mg of sorbent) Waters Sep-Pak Vac Box of 30 C 18 tc 18 (trifunctional) C 8 Silica Florisil Accell Plus CM Accell Plus QMA Alumina A Alumina B Alumina N Amino Propyl (NH 2 ) Cyano Propyl (CN) Diol Part No. WAT020805 WAT036815 WAT036780 WAT052715 WAT020810 WAT020815 WAT020855 WAT020850 WAT020820 WAT020825 WAT020830 WAT020840 WAT020835 WAT020845 Part No. WAT043395 WAT036790 WAT054525 WAT043400 WAT043405 WAT054545 WAT054550 WAT054535 WAT054540 WAT054530 WAT054560 WAT054555 WAT054565 48

Sep-Pak Vac 6 cc Cartridges (contains 1 g of sorbent) Sep-Pak Vac 35 cc Cartridges (contains 10 g of sorbent) Box of 30 Part No. Box of 10 Part No. Waters Sep-Pak Vac C 18 tc 18 (trifunctional) C 8 tc 2 (trifunctional) Silica Florisil Accell Plus CM Accell Plus QMA Alumina A Alumina B Alumina N Amino Propyl (NH 2 ) Cyano Propyl (CN) Diol Sep-Pak Vac 12 cc Cartridges (contains 2 g of sorbent) Waters Sep-Pak Vac Box of 20 C 18 tc 18 (trifunctional) C 8 Silica Florisil Accell Plus CM Accell Plus QMA Alumina A Alumina B Alumina N Amino Propyl (NH 2 ) Cyano Propyl (CN) Diol WAT036905 WAT036795 WAT054570 WAT052705 WAT036910 WAT043390 WAT054590 WAT054595 WAT054580 WAT054585 WAT054575 WAT054605 WAT054600 WAT054610 Part No. WAT036915 WAT043380 WAT054615 WAT036920 WAT043385 WAT054635 WAT054640 WAT054620 WAT054625 WAT054630 WAT054650 WAT054645 WAT054655 Waters Sep-Pak Vac C 18 tc 18 (trifunctional) C 8 Silica Florisil Accell Plus CM Accell Plus QMA Alumina A Alumina B Alumina N Amino Propyl (NH 2 ) Cyano Propyl (CN) Diol Sep-Pak Vac 20 cc Cartridges (contains 5 g of sorbent) Waters Sep-Pak Vac Box of 20 C 18 tc 18 (trifunctional) C 8 Silica Florisil Accell Plus CM Accell Plus QMA Alumina A Alumina B Alumina N Amino Propyl (NH 2 ) Cyano Propyl (CN) Diol WAT043345 WAT043350 WAT054700 WAT043355 WAT043360 WAT054720 WAT054725 WAT054710 WAT054715 WAT054705 WAT054740 WAT054730 WAT054735 Part No. WAT036925 WAT043365 WAT054660 WAT036930 WAT043370 WAT054675 WAT054680 WAT054670 WAT054665 WAT043375 WAT054695 WAT054685 WAT054690 SAMPLE PREPARATION Sep-Pak Cartridge Accessories Description Qty Part No. 30 cc Reservoir 48 WAT011390 60 cc Reservoir 12 WAT024659 Reservoir Adapters for 1, 3 and 6 cc VAC 12 WAT054260 Reservoir Adapters for 12, 20 and 35 cc VAC 10 WAT048160 Male-Male Adapter 100 WAT024310 Male Luer Plugs WAT044395 Female Luer Plugs WAT044385 Sep-Pak Cartridge Connections Kit Contains a selection of the most commonly needed fittings, adapters, valves and tubing for use with Sep-Pak cartridges. Description Sep-Pak Connections Kit Sep-Pak Reservoirs 48/pkg Part No. WAT011400 WAT011390 49

Use Sep-Pak Vac RC cartridges manually or with automated or robotic equipment. The robotic compatible extraction cartridges feature a 20 ml overall capacity which allows for larger conditioning /elution volumes. They eliminate the need for adapting reservoirs to standard size Vac cartridges making sample preparation more convenient. Sep-Pak Vac RC Cartridges are labeled with sorbent type and weight and color-coded for easy identification. Each cartridge contains either 500 mg or 100 mg of high quality sorbent and can be utilized by many robotic sample processors. Sep-Pak Vac RC robotic compatible extraction cartridges contain the same quality and amount of sorbent as our 1 cc - 100 mg and 3 cc - 500 mg Sep-Pak Vac cartridges which makes methods transfer easier. Sep-Pak Vac RC Cartridges (contains 100 mg of sorbent) Sep-Pak Vac RC Cartridges (contains 500 mg of sorbent) Box of 50 Part No. Box of 50 Part No. C 18 tc 18 (trifunctional) C 8 Silica Accell Plus QMA Amino Propyl (NH 2 ) Cyano Propyl (CN) Diol WAT036935 WAT043410 WAT043415 WAT036940 WAT043460 WAT043475 WAT043470 WAT043480 C 18 tc 18 (trifunctional) C 8 Silica Florisil Accell Plus CM Accell Plus QMA Alumina N Amino Propyl (NH 2 ) Cyano Propyl (CN) Diol WAT036945 WAT043425 WAT043430 WAT036950 WAT043435 WAT054505 WAT054500 WAT043485 WAT054515 WAT054510 WAT054520 Read the Primer on SPE - available online. Waters Information Center Visit the most extensive archive of HPLC and SPE applications on the web! Updated daily, this unique web database contains more than 25,000 references to Waters products and their applications. Sources include articles published in refereed journals as well as technical documents and presentations by Waters scientists. click on Applications 50

Sep-Pak DNPH-Silica Cartridges for Analyzing Formaldehyde, Other Aldehydes and Ketones in Air Formaldehyde and other aldehydes are receiving increasing attention both as toxic substances and as promoters in the photochemical formation of ozone in air. Source of aldehydes in residential buildings include plywood and particle board, insulation, combustion appliances, tobacco smoke and various consumer products. Aldehydes are released into the atmosphere in the exhaust of motor vehicles and other equipment in which hydrocarbon fuels are incompletely burned. The most sensitive and specific method for analyzing aldehydes and ketones is based on their reaction with 2,4-dinitrophenylhydrazine (DNPH) and subsequent analysis of the hydrazone derivatives by HPLC. The hydrazones may be detected by absorbance in the ultraviolet region, with maximum sensitivity obtained between 350 and 380 nm. Airborne aldehydes have traditionally been collected by drawing a sample through an impinger containing a solution of DNPH. However, the impinger collector is generally cumbersome to use and is not well suited for high flow rates or extended collection times due to solvent evaporation. The new Sep-Pak DNPH-Silica cartridges meet the requirements of EPA Method TO-11 and provide a convenient device for sample collection. Using a vacuum pump, an air sample is drawn through the new Sep-Pak DNPH-Silica cartridge. The aldehydes and ketones react with the DNPH and form the hydrazone derivative, which is retained on the cartridge. Later, the hydrazones are eluted from the cartridge with acetonitrile and analyzed by HPLC. Detection limits can be as low as 3 ppbv for a 100 liter sample. Advantages of Waters Sep-Pak DNPH-Silica Cartridges HPLC Separation of DNPH Derivatives of Common Aldehydes and Ketones Column: Nova-Pak C 18 3.9 mm x 150 mm Sample: Mixture of DNPH and DNPH derivatives in acetonitrile. Mobile Phase: A: Water/Acetonitrile/Tetrahydrofuran 60/30/10 v/v/v B: Water/Acetonitrile 40/60 v/v 3mL Gradient: 100% A for 1 minute, then linear gradient from 100% A to 100% B in 10 minutes Flow Rate: 1.5 ml/min Injection: 20 µl Detection: 360 nm 1. DNPH 2. Formaldehyde-DNPH 3. Acetaldehyde-DNPH 4. Acetone-DNPH 5. Acrolein-DNPH 6. Propionaldehyde-DNPH 7. Crotonaldehyde-DNPH 8. Butanone-DNPH 9. Butyraldehyde-DNPH 10. Benzaldehyde-DNPH 11. Isovaleraldehyde-DNPH 12. Valeraldehyde-DNPH 13. o-tolualdehyde-dnph 14. m-tolualdehyde-dnph 15. p-tolualdehyde-dnph 16. Hexaldehyde-DNPH 17. 2,5-Dimethylbenzaldehyde-DNPH Low-Level Example: Aldehyde Profile from Laboratory Air Lab Air Sample Cartridge Blank (calculated for 100 liter sample) 1. DNPH 1. DNPH 2. Formaldehyde-DNPH 4.8 ppbv 2. Formaldehyde-DNPH 0.35 ppbv 3. Acetaldehyde-DNPH 1.2 ppbv 3. Acetaldehyde-DNPH 0.27 ppbv 4. Acetone-DNPH 118.0 ppbv 4. Acetone-DNPH 0.34 ppbv 5. Butanone-DNPH 0.8 ppbv 6. Isovaleraldehyde-DNPH 0.7 ppbv SAMPLE PREPARATION These cartridges provide you with significant advantages when compared to other techniques, such as liquid impingers, for the analysis of aldehydes and ketones. In addition, a new high speed, high resolution HPLC application has been developed to provide excellent quantitation capability in the low parts-per-billion range. Sep-Pak DNPH-Silica cartridges meet the requirements of EPA Method TO-11 and ASTM-D-5791-1. Results from impingers and these cartridges are in excellent agreement. Solvent consumption, solvent exposure and hazardous waste disposal costs are reduced. Sep-Pak DNPH-Silica cartridges provide superior convenience and reproducibility, making them ideal for field sampling and process monitoring applications. Sep-Pak DNPH-Silica cartridges can save time and increase productivity. Increased safety. Sep-Pak DNPH-Silica Cartridge Description Quantity Part No. Sep-Pak DNPH-Silica Cartridge Box of 20 WAT037500 Sep-Pak DNPH-Silica Long Body Cartridge Box of 20 WAT039550 The sample of Figure 7 was obtained in a chemical research laboratory using a portable sampling pump. One hundred liters of air was drawn through the cartridge at 0.65 L/min. The sample was found to contain low concentration of formaldehyde (4.8 ppbv), acetaldehyde (1.2 ppbv) and methyl ethyl ketone (0.8 ppbv), but a significant amount of acetone (118 ppbv). Significant concentrations of acetone are frequently found in laboratories because of its widespread use as a solvent. High-Level Example: Aldehyde Profile from Diluted Auto Exhaust Emissions Sample (8.4 L at 1 L/min) collected at US EPA Atmospheric Research Assessment Lab. Courtesy of Dr. S. B. Tejada. 1. DNPH 2. Formaldehyde-DNPH 2290 ppbv 3. Acetaldehyde-DNPH 367 ppbv 4. Acetone-DNPH 26 ppbv 5. Acrolein-DNPH 8 ppbv 6. Propionaldehyde-DNPH 12 ppbv 7. Crotonaldehyde-DNPH 36 ppbv 8. 2-Butanone-DNPH 24 ppbv 9. Butyraldehyde-DNPH 15 ppbv 10. Benzaldehyde-DNPH 32 ppbv 11. o-tolualdehyde-dnph 5 ppbv 12. m-p-tolualdehyde-dnph 11 ppbv 13. 2,5-Dimethylbenzaldehyde-DNPH 3 ppbv 51

Waters XPoSure Aldehyde Sampler Cartridges for Monitoring Aldehydes in indoor air. Based on an extension of our DNPH coating technology, XPoSure Aldehyde Sampler cartridges are the most sensitive active samplers available today. Highest Sensitivity Compared to existing sampling tube technology which have high and variable backgrounds, XPoSure TM cartridges are guaranteed to give consistent low aldehyde backgrounds, cartridge-to-cartridge, lot-to-lot. Low-Level Example: Aldehyde Profile from Laboratory Air 1 2 4 1. DNPH 2. Formaldehyde-DNPH 3. Acetalhyde-DNPH 4. Acetone-DNPH 5. Butanone-DNPH 6. Isovaleraldehyde-DNPH High-Collection Efficiencies You can achieve > 95% collection efficiencies for all aldehydes at flows of up to a liter per minute. And, you only need to use one cartridge - no break- 3 5 6 Lab Air Sample through bed is necessary. Cartridge Blank Low Pressure Drop - Use with Portable Personal Sampling Pumps Large particle size and higher porosity frits make this new XPoSure cartridge compatible with personal sampling pumps. Easy-to-Use Sample, elute and shoot, it s that easy. You ll 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. The figure on the right shows two traces. An actual cartridge blank demonstrating extremely low background levels and as an actual laboratory air sample. 0 2 4 6 8 10 12 14 Time (minutes) The above sample was collected in a chemical research laboratory using a portable sampling pump. One hundred liters of air was drawn through the cartridge at 0.65 L/min. The chromatogram shows ormaldehyde (4.8 ppbv), acetaldehyde (1.2 ppbv), acetone (118 ppbv), butanone (0.8 ppbv), and isovaleraldehyde (0.7 ppbv). XPosure Aldehyde Sampler Cartridge Description Quantity Part No. XPoSure Aldehyde Sampler Cartridge Box of 20 WAT047205 Ozone Scrubber Cartridge Ozone has been shown to interfere with the analysis of carbonyl compounds in air samples that have been drawn through cartridges containing silica-coated with 2,4-dinitrophenylhydrazine (DNPH). Waters Ozone Scrubber cartridges are designed to remove this ozone interference. Flow Schematic for Air Sampling System These disposable devices are intended for use in series combination with the Waters Sep-Pak DNPH-Silica cartridges or XPoSure Aldehyde Sampler cartridges. One Ozone Scrubber cartridge replaces the 1 4 diameter by 36 long copper ozone denuder located in the heated zone of sampling systems used for outdoor air monitoring (PAMS program). Air Sample In O3 Scrubber Cartridge DNPH-Silica or XPoSure Cartridge Vacuum Sampling Pump Each Ozone Scrubber cartridge contains 1.4 grams of granular potassium iodide. When air containing ozone is drawn through this packed bed, iodide is oxidized to iodine, consuming the ozone, according to the following reaction: The theoretical capacity of a single cartridge is 4.2 mmoles of ozone (200 mg). The particle size of the potassium iodide granules is optimized for good mass transfer and flow characteristics. Ozone Scrubber Description Quantity Part No. Ozone Scrubber Box of 20 WAT054420 52

Porapak Rdx Sep-Pak Extraction Cartridge for the Analysis of Explosives in Surface and Ground Waters. Designed to meet or exceed the QA/QC requirements of EPA method 8330, it is ideal for environmental testing laboratories supporting Department of Defense remediation programs. High Sensitivity Activate, Load, Elute and Shoot Flow 15 ml acetonitrile through cartridge at 10 ml/min Flow 30 ml water through cartridge at 10 ml/min Porapak Sep-Pak cartridges contain Porapak Rdx resin, a specially prepared, specially cleaned divinylbenzene/vinylpyrrolidone copolymer, packed in a high purity polypropylene syringe barrel. With the lowest guaranteed backgrounds and the highest cartridge-to-cartridge, lot-to-lot consistency, The Waters Porapak Rdx column is the most sensitive technology available today and allows you to perform analysis at sub ppb levels. Unmatched Recoveries The specially prepared resin is highly selective for nitroaromatic and nitramine compounds, resulting in recoveries of 90% or greater. Recovery data from preconcentrating 500 ml of explosives standards in sterile water at two concentrations on Porapak Rdx Sep-Pak Vac columns. Number of replicates = 7. 1 ppb 10 ppb Compound % Recovery % RSD % Recovery % RSD Load 500 ml water sample at 10 ml/min Elute with 5 ml acetonitrile Dilute to 40% acetonitrile with HPLC grade water Inject 100 µl onto HPLC Isocratic Separation of Method 8330 Analytes Column: Waters Nova-Pak C 8, 3.9 x 150 mm Mobile Phase: 82:18 water/isopropanol Flow rate: 1.0 ml/min Injection volume: 100 ml Temperature: 24 C Detection: UV @ 254 nm Sample: 1 ppb explosives concentrated 40-fold on Porapak Rdx, in 60:40 water/acetonitrile SAMPLE PREPARATION HMX 100.5 6.7 100.5 3.9 TNB 95.9 3.5 99.3 3.3 RDX 90.9 6.4 98.7 3.2 DNB 99.5 3.2 99.2 3.2 TNT 97.0 3.0 102. 3.7 TETRYL 89.0 6.4 102.8 4.7 NB 96.5 2.5 97.9 2.8 3,5-DNA 91.2 3.3 98.2 3.6 2,4-DNT 97.3 3.4 99.9 3.4 2,6-DNT 94.5 3.4 98.7 3.4 2-Am-DNT 92.4 5.2 98.0 3.7 4-Am-DNT 90.0 4.9 97.2 4.1 4-NT 89.5 4.3 100.4 3.7 2-NT 96.8 6.6 93.4 3.0 Increase Productivity and Reduce waste By using Porapak Rdx cartridges, you can reduce the amount of organic solvent used per sample by 10-fold and decrease your sample prep time by 3x. Achieve the lowest method detection limits using Waters Porapak Rdx cartridges. The chromatogram above shows the analysis of reagent H 2 O spiked at the 1 ppb level. The bottom chromatogram is a reagent (H 2 O) blank. Porapak Rdx Cartridges and Accessories Description Quantity Part No. Porapak RDX Cartridges Box of 30 WAT047220 Tubing, Tefzel, 1/8 inch O.D. x 0.040-inch I.D. WAT023344 Sep-Pak Vac Adapter Box of 12 WAT054260 60 cc Sep-Pak Reservoir Box of 12 WAT024659 Male-Male Adapter WAT024310 Sep-Pak Dry SPE Cartridges Waters Sep-Pak Dry cartridges are packed with 2.85 g of anhydrous sodium sulfate. These cartridges are designed to remove residual water from the SPE extract. Sep-Pak Dry Cartridge Description Quantity Part No. Sep-Pak Dry Cartridge Box of 50 WAT054265 53

The Waters Extraction Manifold for Use With Solid-Phase Extraction Cartridges. The vacuum manifold has the capacity to process up to twenty samples simultaneously. The extraction manifold has enhanced features designed for use with conventional silica-based solid phase extraction cartridges as well as modifications that allow you take full advantage of the unique performance characteristics of our Oasis HLB extraction cartridges. B C Just look at what this manifold has to offer: A A B C D Precision machined Delrin cover with alignment posts for quick and easy alignment with test tube rack. Vacuum gauge placement on cover, not in fluid path allows for quick and easy waste removal at bottom by vacuum. Enhanced vacuum control valve designed for use with Waters Oasis HLB extraction cartridges, allows for a quick and momentary rise in vacuum above the frit bubble point at the touch of a finger. High purity polypropylene needle valves and needle tips with minimum dead volume. Extraction Manifolds Description Part No. 20 position extraction manifold without rack (includes 20 needle tips, 25 plugs and ejector tool) WAT200677 20 position extraction manifold with 13 mm x 75 mm test tube rack (includes 20 needle tips, 25 plugs and ejector tool) WAT200606 20 position extraction manifold with 13 mm x 100 mm test tube rack (includes 20 needle tips, 25 plugs and ejector tool) WAT200607 20 position extraction manifold with 16 mm x 75 mm test tube rack (includes 20 needle tips, 25 plugs and ejector tool) WAT200608 20 position extraction manifold with 16 mm x 100 mm test tube rack (includes 20 needle tips, 25 plugs and ejector tool) WAT200609 D: Required when using silica-based SPE cartridges. Accessories and Spare Parts for the Waters Extraction Manifolds Description Part No. Needle valves, pack of 20 (required when using silica-based SPE cartridges) (not required for use with Oasis HLB Extraction Cartridges) WAT200685 Needle tips, pack of 20 WAT200691 Cover, 20 position without gauge assembly WAT200686 Gauge assembly, Vacuum WAT200687 Reservoir, glass with outlet valve WAT200688 Outlet valve kit WAT200689 Gasket for cover WAT200690 Ejector tool WAT058839 Luer Plugs, pack of 25 WAT058851 Rubber ball ring (for vacuum gauge assembly) WAT058840 13 mm x 75 mm test tube rack WAT200678 13 mm x 100 mm test tube rack WAT200679 16 mm x 75 mm test tube rack WAT200680 16 mm x 100 mm test tube rack WAT200681 Reservoir, 30 cc (for Sep-Pak Plus, Light, Vac & Classic cartridges) pkg of 48 WAT011390 Reservoir, 60 cc (for Sep-Pak Plus, Light & Vac cartridges) pkg of 12 WAT024659 Adapter, male-male Luer (for Sep-Pak Classic cartridges) pkg of 100 WAT024310 Adapter (to attach reservoir to 1, 3 & 6 cc Sep-Pak Vac cartridges) pkg of 12 WAT054260 Adapter (to attach reservoir to 12, 20 & 35 cc Sep-Pak Vac cartridges) pkg of 10 WAT048160 Reversible Vial Rack for 1 ml or 4 ml autosampler vials WAT058871 Vacuum pump (110V, 60 Hz) WAT085114 Vacuum pump (220V, 50 Hz) WAT085115 54

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LIMITED PRODUCT WARRANTY Waters Corporation provides this limited warranty (the Warranty) to protect customers from noncomformity in the product workmanship or materials. The Warranty covers all new product manufactured by Waters and its subsidiaries. The Warranty is as follows: Waters warrants that all products sold by them will be of good quality and workmanship. The products will be fit for their intended purposes when used strictly in accordance with Waters instructions for use during the applicable warranty period. The foregoing warranty is exclusive and in lieu of all other express and implied warranties including, but not limited, to fitness for any other purposes(s). In no event will Waters be liable for consequential, economic or incidental damages of any nature. Waters reserves the right not to honor this warranty if the products are abused by the customer. The warranty will not be deemed to have failed of its essential purpose so long as Waters is able and willing to repair or replace any non-conforming part or product. All prices are subject to change without notification. twelve TRADEMARK ACKNOWLEDGEMENTS Trademarks of Waters Corporation: Accell, AccQ Tag, AccQ-Fluor, AccuPure, AccuSep, Alliance, ALPS, Atlantis, Auto Tag, BioSuite, Bondapak, µbondapak, c.news, c.talk, CapLC, C-Gel, Connections, Delta-Pak, DNPH-Silica, EnantioSelect, Envirogel, FractionLynx, Gen-Pak FAX, Guard-Pak, HSPgel, IC-Pak, IonSelect, LC-MALDIprep, LectraBond, M@LDI, Maldi Tof, MassLynx, MassPREP, Micromass, NanoEase, Nova-Pak, Oasis, PIC, Pico Tag, Porapak, Porasil, µporasil, PrepLC, PrepPak, ProteinLynx, Protein-Pak, Q Tof, QuanLynx, Quattro LC, Quattro micro, Quattro Ultima, Radial-Pak, RapiGest, Resolve, SAM, Sentry, Sep-Pak, Silica-Pak, Styragel, µstyragel, Sugar-Pak, Symmetry, Symmetry300, SymmetryPrep, SymmetryShield, Ultrahydrogel, Ultrastyragel, Waters, Waters Spherisorb, XPoSure, XTerra and ZQ Trademarks of YMC, Co., Ltd. Carotenoid, CombiChrom, CombiPrep, CombiScreen, Hydrosphere, J Sphere, N-Sure, ODS-A, ODS-AL, ODS-AM, ODS-AQ, PA-gel, Polyamine II, PolymerC18, Pro C18, Pro C18 RS, Pro C4, Pro C8, Protein-RP, PVA-Sil, R&D Column Set, YMC, YMC*Gel, YMC30, YMCbasic and YMC-Pack. All other trademarks in the catalog are acknowledged. Acrodisc Pall Life Sciences Partisil Whatman AcroPrep Pall Life Sciences Shodex Showa Denko Cheminert VICA Sirocco Pall Life Sciences Florisil US Silica Corporation. Slip-Free Keystone Scientific, Inc. FPLC Pharmacia LKB Supor Pall Life Sciences Inter-Seal Integrated Liner Technologies, Inc. TaqMan Roche Molecular Systems, Inc OptiPak Optimize Technologies Z-Spin Pall Life Sciences µpage J & W ZipTIP Millipore Corporation

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