Maximize HPLC Performance

onsumables olumns and C

Maximize HPLC nalytical HPLC C A Performance

eXtended Performance [XP] 2.5 µm columns allow scientists to improve HPLC throughput and performance compared to 3.5 µm and 5 µm columns while providing a vast selectivity range made possible by 3 unique particles and 14 stationary phases. These columns help maximize productivity and ensure seamless transferability and scalability of LC methods.

118 www.waters.com/order >>> Analytical HPLC Columns AND Consumables

Introduction...... 120 Shodex RSpak Polymer Reversed-Phase Columns...... 149 USP “L” Column Listing...... 122 Paired-Ion Chromatography (PIC Reagents)...... 150 Physical Characteristics of HPLC and UPLC Packing Materials..... 125 Sugar and Carbohydrate Analysis ...... 151 eXtended Performance [XP] 2.5 µm Columns...... 127 Application-Specific Columns...... 154 XSelect Columns...... 129 High-Performance Carbohydrate Analysis Cartridge Column...... 154 XBridge Columns...... 132 Fatty Acid Analysis...... 154 XBridge HILIC Columns for Orthogonal Selectivity...... 135 Pesticide Analysis...... 154 XBridge Amide Columns for Sugar Analysis...... 135 Waters PAH Columns...... 155 SunFire Columns...... 136 Triglycerides and Cholesterol Analysis...... 156 Atlantis Columns...... 138 Fermentation Analysis, Organic Acids, XTerra Columns...... 140 Alcohols, and Carbohydrates...... 156 Symmetry Columns...... 141 Ion Analysis...... 157 Waters Spherisorb Columns...... 144 Anion Analysis...... 157 Waters Spherisorb Analytical Columns...... 144 Cation Analysis...... 157

Waters Spherisorb Cartridge Columns...... 144 Transition Metal Analysis...... 158 A HPLC C HPLC nalytical Nova-Pak Columns...... 145 Ion-Exclusion Columns...... 158 Delta-Pak Columns...... 146 Lanthanide Analysis...... 158 Resolve Columns...... 146 Gas-Chromatography Packings...... 159 µBondapak/Bondapak Packings...... 146 Product Ordering Information...... 160 µPorasil/Porasil Silica Packings...... 146 Guard Products...... 147 Guard-Pak...... 148 C and olumns Waters Sentry Guard Holders and Sentry Guard Columns...... 148 Guard-Pak Holder...... 148 Guard-Pak Inserts...... 148

Literature References onsumables Title Literature Code Title Literature Code Application Notebooks Brochures Atlantis HPLC Columns 720000472EN Atlantis HPLC Columns 720000793EN Symmetry Columns 720000593EN eXtended Performance [XP] 2.5 µm Columns 720004195EN XBridge Amide HPLC Columns 720003438EN HPLC Columns Capabilities 720003995EN XTerra Columns 720000502EN Preparative Optimum Bed Density (OBD) Columns 720002336EN Application Notes SunFire HPLC Columns 720000875EN A New State-of-the-Art Silica C Column 18 WA40526EN Symmetry Columns 720000454EN for Improved Peak Shape of Basic Analytes XBridge HPLC Columns 720001255EN A Quality by Design (QbD) Based Method Development XSelect HPLC Columns 720004178EN for the Determination of Impurities 720004072EN in a Peroxide Degraded Sample of Ziprasidone XTerra Columns 720000424EN A Sensitive Method for the Determination of Endocrine White Papers 720001296EN Disrupting Compounds in River Water by LC/MS/MS A Review of Waters Hybrid Particle Technology Determination of Soy Isoflavones in Part 2. Ethylene-Bridged (BEH Technology) Hybrids and 720001159EN 720004253EN Foods and Dietary Supplements by UPLC Their Use in Liquid Chromatography 720004098EN Charged Surface Hybrid (CSH) Technology Extract using Charged Surface Hybrid (CSH) Technology 720003469EN Modernization of the USP Assay for Clarithromycin Using and Its Use in Liquid Chromatography 720004461EN eXtended Performance [XP] Column Technology Topics in Liquid Chromatography: Practical Applications of Charged Part 1. Designing a Reversed-Phase Column 720001889EN 720003720EN Surface Hybrid (CSH) Technology for Polar Compound Retention Rapid, Reliable Metabolite Ratio Evaluation for MIST Online Tools 720004453EN Assessments in Drug Discovery and Preclinical Studies Waters Column Advisor www.waters.com/columnadvisor Utlizing XBridge HPLC Columns for Waters Reversed-Phase WA43181 www.waters.com/selectivitychart Method Development at pH Extremes Column Selectivity Chart Validation of a Method for the Separation of Ziprasidone Books and Its Degradants Using Empower 2 with Method 720004077EN Beginner’s Guide to Liquid Chromatography 715001531 Validation Manager (MVM) Comprehensive Guide to HILIC 715002531 HPLC Columns–Theory, Technology, and Practice WAT038216 Search by Literature Code at waters.com 119 Dependable HPLC Column Performance

Choosing an HPLC column can be a very difficult process. The chromatographic packing material (particle ‘brand name’ and functional group) plays a major role in the success of a separation, both initially, when the method is first developed, as well as long term, if the method will need to be reproduced for many years. onsumables olumns and C

HPLC Columns: Theory, Technology, and Practice nalytical HPLC C Uwe D. Neue, Waters Corporation, Milford, MA, published by John Wiley & Sons A High-performance liquid chromatography and its derivative techniques have become the dominant analytical separation tools in the pharmaceutical, chemical, and food industries, environmental laboratories, and therapeutic drug monitoring. Although the column is the heart of the HPLC instrument and essential to its success, until now no single book has focused on the theory and practice of column technology. ‘HPLC Columns’ provides thorough, state-of-the-art coverage of HPLC column technology for the practicing technician and academician alike. Along with a comprehensive discussion of the chemical and physical processes of the HPLC column, it includes fundamental principles, separation mechanisms, available technologies, column selection criteria, and special techniques.

Special features include: Explanation of the underlying principles of HPLC columns Methods for selecting columns Practical advice on using and applying columns, including examples Section by M. Zoubair El Fallah on methods development Description Part No. HPLC Columns: Special techniques, including preparative chromatography, Theory, Technology, WAT038216 continuous chromatography, and the simulated moving bed and Practice Troubleshooting

120 www.waters.com/order USP “L” Column Listing

US Pharmacopeia LC “L1” (C18) Column Designation Subclassification Testing by Brand Name*

Hydrophobicity Chelating Silanol Activity Silanol Activity Shape Selectivity Column Brand Name (k, Ethylbenzene) (Tailing Factor [TF], Quinizarin) (k, Amitriptyline) (TF, Amitriptyline) (Bonding Density, µmol/m2)

ACQUITY UPLC BEH C18 1.63 1.10 3.79 1.10 3.10 ACQUITY UPLC BEH Shield RP18 1.30 1.36 2.37 1.13 3.30

ACQUITY UPLC HSS C18 2.54 1.36 5.96 1.41 3.20

ACQUITY UPLC HSS C18 SB 1.09 1.07 22.20 7.20 1.60 ACQUITY UPLC HSS T3 2.23 1.19 6.50 3.15 1.60

Atlantis dC18 1.70 1.10 5.10 2.40 1.60 Atlantis T3 1.87 0.98 5.32 1.35 1.60

™ Delta-Pak C18 (100Å) 2.00 2.00 6.30 1.90 3.60

® µBondapak C18 1.00 6.00 7.50 4.00 1.10

Nova-Pak C18 1.50 7.50 4.60 3.00 2.70

™ Resolve C18 2.00 no peak 35.0 8.00 2.80

SunFire C18 2.50 1.20 6.30 1.10 3.50 A HPLC C HPLC nalytical Symmetry C18 2.25 1.70 5.10 1.70 3.20 SymmetryShield RP18 1.65 1.50 3.10 1.20 3.30 Waters Spherisorb® ODS-1 0.80 no peak 23.00 3.00 1.70 Waters Spherisorb ODS-2 2.00 no peak 11.50 7.00 2.60

XBridge C18 1.63 1.10 3.79 1.10 3.10 XBridge Shield RP18 1.30 1.36 2.37 1.13 3.30

XSelect HSS C18 2.54 1.36 5.96 1.41 3.20 C and olumns

XSelect HSS C18 SB 1.09 1.07 22.20 7.20 1.60 XSelect HSS T3 2.23 1.19 6.50 3.15 1.60 XTerra RP18 1.00 1.20 1.70 1.10 2.30

XTerra MS C18 1.50 1.10 3.30 1.30 2.20

* For use in C18 column brand name selection when referencing USP monographs which specify the characteristics/performance of the C18 column used in the method. Choose a brand as close as possible to the referenced column values for the best chance of duplicating the analytical results of the method. onsumables

Waters Electronic Column Selectivity Chart

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121 USP “L” Column Listing L3 Porous silica particles–1.5 to 10 µm in diameter. Brand Particle Size Type Page Octadecylsilane (ODS or C18) chemically bonded to porous L1 silica or ceramic particles–1.5 to 10 µm in diameter. ACQUITY UPLC BEH HILIC 1.7 µm Spherical 99 See the subclassification table on previous page. Atlantis HILIC Silica 3, 5 µm Spherical 138 Brand Particle Size Type Page BioSuite UHR SEC 5, 8 µm Spherical 241 AccQ•Tag™ Ultra 1.7 µm Spherical 247 BioSuite SEC 7.5 µm Spherical 241 Nova-Pak 6, 4 µm Spherical 145 ACQUITY UPLC BEH C18 1.7 µm Spherical 99 ACQUITY UPLC BEH Shield RP18 1.7 µm Spherical 99 µPorasil 10 µm Spherical 146 Resolve 5, 10 µm Spherical 146 ACQUITY UPLC CSH C18 1.7 µm Spherical 97 SunFire Silica 5, 10 µm Spherical 136 ACQUITY UPLC HSS C18 1.7 µm Spherical 101 Waters Spherisorb 5, 10 µm Spherical 144 ACQUITY UPLC HSS C18 SB 1.8 µm Spherical 101 ACQUITY UPLC HSS T3 1.8 µm Spherical 101 XBridge HILIC 2.5, 3.5, 5 µm Spherical 132

ACQUITY UPLC OST C18 1.7 µm Spherical 251 Atlantis T3 3, 5, 10 µm Spherical 138 Silica gel of a controlled surface porosity bonded to a solid

Atlantis dC18 3, 5, 10 µm Spherical 138 L4 spherical core–30 to 50 µm in diameter. BioSuite™ PA-A 3 µm Spherical 225 Brand Particle Size Type Page BioSuite PA-B 3 µm Spherical 225 Porasil™ Prep Silica 50 µm Irregular 146 Delta-Pak C18 5 µm Spherical 146

µBondapak C18 10 µm Irregular 146 ™ µBondapak C18 Radial-Pak 10 µm Irregular 146 Alumina of controlled surface porosity bonded to a solid spherical core–30 to 50 µm in diameter. nanoACQUITY UPLC BEH130 1.7 µm Spherical 261 L5 nanoACQUITY UPLC BEH300 1.7 µm Spherical 261

Nova-Pak C18 4, 6 µm Spherical 145 Strong cation exchanger packing-sulfonated fluorocarbon polymer

Prep Nova-Pak HR C18 6 µm Spherical 194 L6 coated on a solid spherical core–30 to 50 µm in diameter.

Resolve C18 5, 10 µm Spherical 146 onsumables

SunFire C18 2.5, 3.5, 5, 10 µm Spherical 136 Octyl silane (C8) chemically bonded to porous silica particles– Symmetry C18 3.5, 5, 7 µm Spherical 141 L7 1.5 to 10 µm in diameter. ™ SymmetryPrep C18 3.5, 5, 7 µm Spherical 194 ™ Brand Particle Size Type Page Symmetry300 C18 3.5, 5 µm Spherical 194 ACQUITY UPLC BEH C 1.7 µm Spherical 99 SymmetryShield RP18 3.5, 5 µm Spherical 141 8 Nova-Pak C 4, 6 µm Spherical 145 Waters Spherisorb ODS1 3.5, 5, 10 µm Spherical 144 8 146 olumns and C Resolve C 5, 10 µm Spherical Waters Spherisorb ODS2 3.5, 5, 10 µm Spherical 144 8 Waters Spherisorb C 3, 5, 10 µm Spherical 144 Waters Spherisorb ODSB 3.5, 5, 10 µm Spherical 144 8 SunFire C 3.5, 5, 10 µm Spherical 136 XBridge BEH130 3.5, 5, 10 µm Spherical 132 8 Symmetry C 3.5, 5, 7 µm Spherical 141 XBridge BEH300 3.5, 5, 10 µm Spherical 132 8 SymmetryShield RP8 3.5, 5 µm Spherical 141 XBridge C18 2.5, 3.5, 5, 10 µm Spherical 132 SymmetryPrep C8 7 µm Spherical 194 XBridge OST C18 2.5 µm Spherical 251 XBridge C 2.5, 3.5, 5, 10 µm Spherical 132 XBridge Shield RP18 2.5, 3.5, 5, 10 µm Spherical 132 8 XTerra MS C8 2.5, 3.5, 5, 10 µm Spherical 140 XSelect CSH C18 2.5, 3.5, 5 µm Spherical 129 XTerra RP8 3.5, 5, 10 µm Spherical 140 nalytical HPLC C XSelect HSS C18 2.5, 3.5, 5 µm Spherical 129 A XSelect HSS C18 SB 2.5, 3.5, 5 µm Spherical 129 XSelect HSS T3 2.5, 3.5, 5 µm Spherical 129 An essentially monomolecular layer of aminopropylsilane

XTerra MS C18 2.5, 3.5, 5, 10 µm Spherical 140 (NH2) chemically bonded to totally porous silica gel support– L8 3 to 10 µm in diameter. XTerra RP18 3.5, 5, 10 µm Spherical 140 Brand Particle Size Type Page 146 Octadecylsilane (ODS or C18) chemically bonded to silica gel µBondapak NH2 10 µm Irregular of a controlled surface porosity bonded to a solid spherical High Performance Carbohydrate 3, 5 µm — 152 L2 core–30 to 50 µm in diameter. Analysis Waters Spherisorb NH 3, 5, 10 µm Spherical 144 Brand Particle Size Type Page 2

® Bondapak Prep C18 50 µm Irregular 146 3 to 10 µm irregular, totally porous silica gel having a chemically L9 bonded strongly acidic cation exchanger coating (SCX). Brand Particle Size Type Page Waters Spherisorb SCX 5, 10 µm Spherical 144

( ) - Denotes particle sizes available outside of L class.

122 www.waters.com/order Nitrile groups (CN) chemically bonded to porous silica particles– Amino (NH2) and Cyano (CN) groups chemically bonded to L10 3 to 10 µm in diameter. L18 porous silica particles–3 to 10 µm in diameter. Brand Particle Size Type Page ACQUITY UPLC HSS CN 1.8 μm Spherical 101 Strong cation exchange resin consisting of sulfonated, cross-linked styrene divinylbenzene copolymer in the calcium µBondapak CN 10 µm Irregular 146 L19 form–about 9 µm in diameter. Nova-Pak CN HP 4 µm Spherical 145 Resolve CN 5, 10 µm Spherical 146 Brand Particle Size Type Page Waters Spherisorb CN 3, 5, 10 µm Spherical 144 Sugar-Pak™ 1 9 µm Spherical 178 XSelect HSS CN 2.5, 3.5, 5 μm Spherical 129 Shodex SC-1011 7 µm Spherical 243

Phenyl groups chemically bonded to porous silica particles– Dihydroxypropane groups chemically bonded to porous silica L11 1.5 to 10 µm in diameter. L20 particles–3 to 10 µm in diameter. Brand Particle Size Type Page Brand Particle Size Type Page 97 4, 5, 8, 10, ACQUITY UPLC CSH Phenyl-Hexyl 1.7 µm Spherical BioSuite 125, 250, 450 Spherical 241 ACQUITY UPLC BEH Phenyl 1.7 µm Spherical 99 13, 17 µm µBondapak Phenyl 10 µm Irregular 146 Insulin HMWP — N /A 226 ™ Nova-Pak Phenyl 4 µm Spherical 145 Protein-Pak 60 10 µm Irregular 243 XBridge Phenyl 2.5, 3.5, 5 µm Spherical 132 Protein Pak 125 10 µm Irregular 243 Waters Spherisorb Phenyl 3, 5, 10 µm Spherical 144 Protein-Pak 300SW 10 µm Irregular 243 A XSelect CSH Phenyl-Hexyl 2.5, 3.5, 5 µm Spherical 129 Protein-Pak KW-802.5 7 µm Irregular 272 HPLC C HPLC nalytical XTerra Phenyl 3.5, 5 µm Spherical 140 Protein-Pak KW-803 7 µm Irregular 272 Protein-Pak KW-804 7 µm Irregular 272

A strong anion exchanger packing made by chemically bonding a quaternary amine to a solid silica spherical core–30 to 50 µm A rigid, spherical styrene-divinylbenzene copolymer 5 to 10 µm L12 in diameter. L21 in diameter.

Brand Particle Size Type Page Brand Particle Size Type Page Accell Plus QMA 50 µm Irregular 239 Shodex RSpak 613 6 µm Spherical 149

Styragel® HR 0.5, 1, 2, 3, and 4 — Spherical 282 C and olumns Styragel HR 4E — Spherical 282 Trimethylsilane (C1) chemically bonded to porous silica particles– L13 3 to 10 µm in diameter. Styragel 5E — Spherical 282 Brand Particle Size Type Page A cation-exchange resin made of porous polystyrene with Waters Spherisorb C 3, 5, 10 µm Spherical 144 1 L22 sulfonic acid groups–about 10 µm in size.

Brand Particle Size Type Page onsumables Silica gel having a chemically bonded, strongly basic quaternary L14 ammonium anion exchanger (SAX) coating–5 to 10 µm in diameter. IC-Pak Ion Exclusion 7 Spherical 158 Shodex RSpak DC 613 6 Spherical 149 Brand Particle Size Type Page Shodex SP-0810 8 Spherical 243 Waters Spherisorb SAX 5, 10 µm Spherical 144

An anion-exchange resin made of porous polymethacrylate Hexylsilane (C6) chemically bonded to a totally porous silica L23 or polyacrylate gel with quaternary ammonium groups–about L15 particle–3 to 10 µm in diameter. 10 µm in size. Brand Particle Size Type Page Brand Particle Size Type Page

Waters Spherisorb C6 3, 5, 10 µm Spherical 144 BioSuite Q AXC 10, 13 µm Spherical 237 BioSuite DEAE 2.5, 10, 13 µm Spherical 237 BioSuite Q-PEEK 10 µm Spherical 237 Dimethylsilane (C2) chemically bonded to a totally porous L16 silica particles–5 to 10 µm in diameter. IC-Pak Anion 10 µm Spherical 158 Protein-Pak Q 8HR 8 µm Spherical 243

Strong cation exchange resin consisting of sulfonated, cross-linked styrene divinylbenzene copolymer in the hydrogen A semi-rigid hydrophilic gel consisting of vinyl polymers with L17 form–7 to 11 µm in diameter. numerous hydroxyl groups on the matrix surface–32 to 63 µm L24 in diameter. Brand Particle Size Type Page Fast Fruit Juice N /A N /A 178 IC-Pak™ Ion Exclusion 7 µm Spherical 158 IC-Pak Cation 10 µm Irregular 158 Shodex® RSpak DC-613 (6) µm Spherical 149

( ) - Denotes particle sizes available outside of L class.

123 Packing having the capacity to separate compounds with a 3,5-dinitrobenzoyl derivative of L-phenylglycine covalently molecular weight range from 100 to 5,000 (as determined by L36 bonded to a 5 µm aminopropyl silica. polyethylene oxide), applied to neutral, anionic and cationic L25 water-soluble polymers. A polymethacrylate resin base, cross- linked with polyhydroxylated ether (surface contained some Packing having the capacity to separate proteins by molecular residual carboxyl groups), was found suitable. size over a range of 2,000 to 40,000 daltons. It is a L37 polymethacrylate gel. Brand Particle Size Type Page Brand Particle Size Type Page Ultrahydrogel™ DP, + 120 10 µm Spherical 282 Ultrahydrogel 250 N /A Spherical 289

Butyl silane (C4) chemically bonded to porous silica particles– L26 3 to 10 µm in diameter. A methacrylate-based size-exclusion packing for water L38 soluble samples. Brand Particle Size Type Page Brand Particle Size Type Page ACQUITY UPLC BEH300 C4 1.7 µm Spherical 99 Ultrahydrogel N /A Spherical 289 Spherical Delta-Pak C 5 µm 146 4 (100 + 300Å) A hydrophilic-polyhydroxymethacrylate gel of totally porous Symmetry300 C4 3.5 µm Spherical 244 L39 spherical resin. XBridge BEH300 C4 3.5 µm Spherical 132 Brand Particle Size Type Page L27 Porous silica particles–30 to 50 µm in diameter. Ultrahydrogel N /A Spherical 289 Brand Particle Size Type Page Cellulose tris-3,5-dimethylphenylcarbamate coated porous Porasil 37–55 µm Irregular 146 L40 silica particles–5 to 20 µm in diameter.

A multifunctional support which consists of a high purity, Immobilized a1-acid glycoprotein on spherical silica particles– 100Å, spherical silica substrate that has been bonded with L41 5 µm in diameter. L28 anionic (amine) functionality in addition to a conventional onsumables reversed-phase C8 functionality. Octylsilane and octadecylsilane groups chemically bonded to L42 porous silica particles–5 µm in diameter. Gamma alumina, reversed-phase, low carbon percentage by L29 weight alumina-based polybutadiene spherical particles–5 µm Pentafluorophenyl groups chemically bonded to silica particles– in diameter with a pore diameter of 80Å. L43 5 to 10 µm in diameter. Brand Particle Size Type Page Ethyl silane chemically bonded to a totally porous silica olumns and C L30 particle–3 to 10 µm in diameter. ACQUITY UPLC CSH Fluoro-Phenyl 1.7 µm Spherical 97 ACQUITY UPLC HSS PFP 1.8 µm Spherical 101 A strong anion-exchange resin-quaternary amine bonded on XSelect CSH Fluoro-Phenyl 2.5, 3.5, 5 µm Spherical 129 latex particles attached to a core of 8.5 µm macroporous particles XSelect HSS PFP 2.5, 3.5, 5 µm Spherical 129 L31 having a pore size of 2,000Å and consisting of ethylvinylben- zene cross-linked with 55% divinyl benzene. A multifunctional support, which consists of a high purity, 60Å, spherical silica substrate that has been bonded with a cationic A chiral-ligand exchange packing–L proline copper complex L44 exchanger, sulfonic acid functionality in addition to a convention nalytical HPLC C covalently bonded to irregularly shaped silica particles–5 to reversed-phase C8 functionality. A L32 10 µm in diameter.

Beta cyclodextrin bonded to porous silica particles–5 to 10 µm Packing having the capacity to separate proteins of 4,000 to L45 in diameter. 400,000 daltons. It is spherical, silica-based and processed L33 to provide pH stability. Polystyrene/divinylbenzene substrate agglomerated with quaternary Brand Particle Size Type Page L46 amine functionalized latex beads–10 µm in diameter. BEH125 SEC 1.7 µm Spherical 227 BEH200 SEC 1.7 µm Spherical 227 A strong cation-exchange resin made of porous silica coated with L55 polybutadiene-maleic acid copolymer–about 5 µm in diameter. Strong cation-exchange resin consisting of sulfonated cross- Brand Particle Size Type Page linked styrene-divinylbenzene copolymer in the lead form– IC-Pak C M/D — — 148 L34 about 9 µm in diameter.

Brand Particle Size Type Page Packing having the capacity to separate proteins by molecular Shodex SP-0810 N /A Spherical 243 weight over the range of 10 to 500 kDa. It is spherical (10 µm), L59 silica-based, and processed to provide hydrophilic characteristics and pH stability. Zirconium-stabilized spherical silica packing with a hydrophilic (diol-type) molecular mono layer bonded phase having a pore Brand Particle Size Type Page L35 size of 150Å. BioSuite 125, 250, 450 Series 4–17 µm Spherical 241

Source: United States Pharmacopeia ( ) - Denotes particle sizes available outside of L class.

124 www.waters.com/order Physical Characteristics of HPLC and UPLC Packing Materials

Pore Surface Area Pore Volume % Carbon Brand Chemistry Particle Shape Particle Size(s) End Capped Size* [m2/g] [cc/g] Load

AccQ•Tag Ultra C18 Spherical 1.7 µm 135Å 185 0.70 17 Ye s

C18 Spherical 1.7 µm 130Å 185 0.70 17 Ye s

C8 Spherical 1.7 µm 130Å 185 0.70 13 Ye s Shield RP18 Spherical 1.7 µm 130Å 185 0.70 17 Ye s ACQUITY UPLC BEH Phenyl Spherical 1.7 µm 130Å 185 0.70 15 Ye s HILIC Spherical 1.7 µm 130Å 185 0.70 Unbonded n/a Amide Spherical 1.7 µm 130Å 185 0.70 12 n/a ACQUITY UPLC BEH125 SEC Spherical 1.7 µm 125Å 398 High 15 n/a

ACQUITY UPLC BEH130 C18 Spherical 1.7 µm 130Å 185 0.70 17 Ye s ACQUITY UPLC BEH200 SEC Spherical 1.7 µm 200Å 220 High 12 n/a

C18 Spherical 1.7 µm 300Å 90 0.66 12 Ye s ACQUITY UPLC BEH300 C4 Spherical 1.7 µm 300Å 90 0.66 8 No

C18 Spherical 1.7 µm 130Å 185 0.70 15 Ye s Phenyl-Hexyl Spherical 1.7 µm 130Å 185 0.70 14 Ye s ACQUITY UPLC CSH Fluoro-

Spherical 1.7 µm 130Å 185 0.70 10 No A Phenyl HPLC C HPLC nalytical T3 Spherical 1.8 µm 100Å 230 0.70 11 Ye s

C18 Spherical 1.8 µm 100Å 230 0.70 15 Ye s

ACQUITY UPLC HSS C18 SB Spherical 1.8 µm 100Å 230 0.70 8 No PFP Spherical 1.8 µm 100Å 230 0.70 7 No CN Spherical 1.8 µm 100Å 230 0.70 5 No T3 Spherical 3, 5, 10 µm 100Å 330 1.00 14 Ye s

Atlantis dC18 Spherical 3, 5, 10 µm 100Å 330 1.00 12 Ye s C and olumns HILIC Spherical 3, 5 µm 100Å 330 1.00 Unbonded n/a

C18 Irregular 15–20 µm 125Å 330 1.00 10 Ye s Bondapak C18 Irregular 15–20 µm 300Å 100 1.00 3.5 Ye s

C18 Irregular 10 µm 125Å 330 1.00 9.8 Ye s Phenyl Irregular 10 µm 125Å 330 1.00 9.3 Ye s µBondapak CN Irregular 10 µm 125Å 330 1.00 6 Ye s onsumables

NH2 Irregular 10 µm 125Å 330 1.00 4 No

C4 Spherical 5, 15 µm 100Å 300 1.00 7.3 Ye s

C18 Spherical 5, 15 µm 100Å 300 1.00 17 Ye s Delta-Pak C4 Spherical 5, 15 µm 300Å 125 1.00 2.6 Ye s

C18 Spherical 5, 15 µm 300Å 125 1.00 6.8 Ye s

C18 Spherical 4, 6 µm 60Å 120 0.30 7.3 Ye s

C8 Spherical 4 µm 60Å 120 0.30 4 Ye s Nova-Pak Phenyl Spherical 4 µm 60Å 120 0.30 4.6 Ye s CN HP Spherical 4 µm 60Å 120 0.30 3 Ye s Silica Spherical 4, 6 µm 60Å 120 0.30 n/a n/a Porasil Silica Irregular 15–20 µm 125Å 330 1.00 n/a n/a µPorasil™ Silica Irregular 10 µm 125Å 330 1.00 n/a n/a

C18 Spherical 5, 10 µm 90Å 200 0.50 10.2 No

C8 Spherical 5 µm 90Å 200 0.50 5.1 No Resolve CN Spherical 10 µm 90Å 200 0.50 3 No Silica Spherical 5, 10 µm 90Å 200 0.50 n/a n/a

* Nominal value

125 Physical Characteristics of HPLC and UPLC Packing Materials

Pore Surface Area Pore Volume % Carbon Brand Chemistry Particle Shape Particle Size(s) End Capped Size* [m2/g] [cc/g] Load

C18 Spherical 2.5, 3.5, 5, 10 µm 100Å 340 0.90 16 Ye s

SunFire C8 Spherical 2.5, 3.5, 5, 10 µm 100Å 340 0.90 12 Ye s Silica Spherical 5, 10 µm 100Å 340 0.90 Unbonded n/a

C18 Spherical 3.5, 5 µm 100Å 335 0.90 19.1 Ye s Symmetry C8 Spherical 3.5, 5 µm 100Å 335 0.90 11.7 Ye s

C18 Spherical 3.5, 5 µm 300Å 110 0.80 8.5 Ye s Symmetry300 C4 Spherical 3.5, 5 µm 300Å 110 0.80 2.8 Ye s

C18 Spherical 7 µm 100Å 335 0.90 19.1 Ye s SymmetryPrep C8 Spherical 7 µm 100Å 335 0.90 11.7 Ye s RP8 Spherical 3.5, 5 µm 100Å 335 0.90 15 Ye s SymmetryShield RP18 Spherical 5 µm 100Å 335 0.90 17 Ye s Silica Spherical 3, 5, 10 µm 80Å 220 0.50 n/a n/a ODS2 Spherical 3, 5, 10 µm 80Å 220 0.50 11.5 Ye s ODS Spherical 3, 5, 10 µm 80Å 220 0.50 6.2 No ODSB Spherical 5 µm 80Å 220 0.50 11.5 Ye s

C8 Spherical 3, 5, 10 µm 80Å 220 0.50 5.8 Ye s

C6 Spherical 3, 5, 10 µm 80Å 220 0.50 4.7 Ye s Waters Spherisorb C1 Spherical 3, 5, 10 µm 80Å 220 0.50 2.2 No Nitrile Spherical 3, 5, 10 µm 80Å 220 0.50 3.1 No Amino Spherical 3, 5, 10 µm 80Å 220 0.50 1.9 No onsumables Phenyl Spherical 3, 5, 10 µm 80Å 220 0.50 2.5 No OD/CN Spherical 5 µm 80Å 220 0.50 5 Ye s SAX, SCX Spherical 5, 10 µm 80Å 220 0.50 4 No

C18 Spherical 2.5, 3.5, 5, 10 µm 130Å 185 0.70 18 Ye s

C8 Spherical 2.5, 3.5, 5, 10 µm 130Å 185 0.70 13 Ye s Shield RP18 Spherical 2.5, 3.5, 5, 10 µm 130Å 185 0.70 17 Ye s XBridge olumns and C Phenyl Spherical 2.5, 3.5, 5 µm 130Å 185 0.70 15 Ye s HILIC Spherical 2.5, 3.5, 5 µm 130Å 185 0.70 Unbonded n/a Amide Spherical 2.5, 3.5 µm 130Å 185 0.70 12 n/a

XBridge BEH130 C18 Spherical 3.5, 5, 10 µm 130Å 185 0.70 18 Ye s

C18 Spherical 3.5, 5, 10 µm 300Å 86 0.66 12 Ye s XBridge BEH300 C4 Spherical 3.5 µm 300Å 90 0.66 8 No

C18 Spherical 2.5, 3.5, 5 µm 130Å 185 0.70 15 Ye s nalytical HPLC C XSelect CSH Phenyl-Hexyl Spherical 2.5, 3.5, 5 µm 130Å 185 0.70 14 Ye s A Fluoro-Phenyl Spherical 2.5, 3.5, 5 µm 130Å 185 0.70 10 No T3 Spherical 2.5, 3.5, 5 µm 100Å 230 0.70 11 Ye s

C18 Spherical 2.5, 3.5, 5 µm 100Å 230 0.70 15 Ye s

XSelect HSS C18 SB Spherical 2.5, 3.5, 5 µm 100Å 230 0.70 8 No PFP Spherical 2.5, 3.5, 5 µm 100Å 230 0.70 7 No CN Spherical 2.5, 3.5, 5 µm 100Å 230 0.70 5 No RP18 Spherical 3.5, 5, 10 µm 125Å 175 0.70 15 Ye s RP8 Spherical 3.5, 5, 10 µm 125Å 175 0.70 13.5 Ye s

XTerra MS C18 Spherical 2.5, 3.5, 5, 10 µm 125Å 175 0.70 15.5 Ye s

MS C8 Spherical 2.5, 3.5, 5, 10 µm 125Å 175 0.70 12 Ye s Phenyl Spherical 3.5, 5 µm 125Å 175 0.70 12 Ye s * Nominal value

126 www.waters.com/order eXtended Performance [XP] 2.5 µm Columns

XBridge and XSelect eXtended Performance [XP] 2.5 μm columns enable exceptional separation performance, robustness, and throughput for HPLC assays while being fully compatible with all HPLC, UHPLC, and UPLC technology platforms. Unmatched Selectivity and Flexibility.

14 chemistries including C18, Phenyl-Hexyl, C8, Embedded Polar, HILIC, Amide as well as Cyano and PFP columns Directly scalable to 1.7/1.8 μm UPLC columns and larger 3.5/5 μm HPLC columns Designed to withstand higher pressures of 9,000 psi (4.6 mm ID) and 18,000 psi (2.1 and 3.0 mm ID) Select column length (30, 50, 75, 100, and 150 mm) for the correct balance between resolution and throughput

High Throughput. Lower Backpressure. A Improved Productivity.  HPLC C HPLC nalytical 40% lower backpressure than sub-2-μm UPLC columns See page 170 Equivalent backpressure and superior performance to for part numbers and core-shell columns ordering information. 2–4X higher throughput than 3.5–5-μm HPLC columns lead to improved productivity Rapid Method Optimization. C and olumns Transfer methods TODAY within current pharmacopeia guidelines Improve Existing HPLC Productivity 2–4X Utilize on an ACQUITY UPLC system to maximize performance and future-proof your method 5

XSelect CSH C18 4.6 x 100 mm, 3.5 µm 1 2 Part Number: 186005269 HPLC System onsumables Did you know... 3 You can request Certificates of Analysis on our website? 4 Go to: www.waters.com and look for “Request Certificate of 0.0 4.0 8.0 12.0 16.0 20.0 24.0 28.0 min Analysis” under “Services and Support”

XSelect CSH C18 XP 4.6 x 75 mm, 2.5 µm Part Number: 186006110 HPLC System

2X Faster than 3.5 µm HPLC

0.0 2.0 4.0 6.0 8.0 10.0 12.0 14.0 min

ACQUITY UPLC CSH C18 2.1 x 50 mm, 1.7 µm Part Number: 186005296 UPLC System

9X Faster than 3.5 µm HPLC

5X Faster than 2.5 µm HPLC

0.0 0.5 1.0 1.5 2.1 2.5 3.0 min

Peak Order: (1) tolmetin, (2) naproxen, (3) fenoprofen, (4) indomethacin, (5) dicolfenact

127 Superior Performance vs. Core-Shell with Equivalent Backpressure XP 2.5 µm columns provide 34% higher peak capacity than equivalent core-shell columns due to the limited loading capacity of this type of particle technology. Additionally, both columns exhibit similar backpressures indicating that the particle size dictates the observed backpressure, not the particle type (core-shell vs. fully-porous).

Equivalent Backpressure 34% Higher Peak Capacity

8000 XSelect CSH C18 XP 0.30 XBridge BEH C18 XP, 2.5 µm 4.6 x 75 mm, 2.5 µm

7000 Pindolol 0.25 Part Number: 186006110 Core-Shell C18,2.6 µm Peak Capacity = 47 Diltazem 6000 0.20

5000 0.15 Quinine

4000 0.10 Verapamil Labetolol

Pressure (psi) 3000 0.05

0.00 2000 0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 min 1000 2.1 x 50 mm columns 70/30 MeCN/H 0, 30 °C, 254 nm Core-Shell C18 2 0.30 0 4.6 x 75 mm, 2.6 µm 0.0 0.51.0 1.52.0 2.5 0.25 Peak Capacity = 35 Flow Rate (mL/min) 0.20 Comparative separations may not be representative of all applications. 0.15

0.10

0.05 onsumables 0.00

0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 min

Comparative separations may not be representative of all applications. olumns and C Additional Flexibility for UPLC Users XP 2.5 µm columns provide an alternative to 1.7 µm UPLC columns for use on an ultra-low dispersion ACQUITY UPLC instrument. With 40% lower backpressure than 1.7 µm columns, XP 2.5 µm columns can be utilized for fit-for-purpose analyses at intermediate backpressures. Alternatively, flow rate can be increased to further expedite sample throughput for simple chromatography.

40% Lower Backpressure on Your UPLC nalytical HPLC C

A ACQUITY UPLC CSH C18 XSelect CSH C18 XP 0.35 R = 6.3 0.35 S 2.1 x 75 mm, 1.7 µm 2.1 x 75 mm, 2.5 µm Part Number: 186005620 0.30 Part Number: 186006102 0.30 R = 5.5 Flow Rate = 0.54 mL/min S Flow Rate = 0.54 mL/min 0.25 0.25 Pressure = 9200 PSI Pressure = 5300 PSI 0.20 0.20 Diclofenac Peak Capacity = 40 Peak Capacity = 34 AU AU 0.15 0.15

Tolmetin 15% increase in Rs and Pc 40% lower 0.10 20% higher sensitivity 0.10 Naproxen backpressure Fenoprofen 0.05 0.05 Indomethacin 0.00 0.00 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 min 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 min

Columns were run on an ACQUITY UPLC H-Class system.

128 www.waters.com/order XSelect Columns XSelect columns offer the separations scientist the most diverse range of modern LC columns for method development. The scientists that use XSelect columns demand diverse selectivity and realize that it is the combination of base particle composition and ligand bonding that critically influences analyte selectivity and column performance. It is the optimized combination of substrate and ligand that creates a column family for enhancing analyte selectivity. The XSelect column family is offered in two distinct substrates (High Strength Silica [HSS] and Charged Surface Hybrid [CSH]) to give you the most option

and control over your separation power. CSH Technology Advantages  Superior peak shape for basic compounds See page 172 - High peak capacity for complex mixtures for part numbers and ordering information. Increased loading capacity - Increased mass load for preparative separation without using ion-pairing reagents like TFA A

Rapid column equilibration C HPLC nalytical - Quickly change mobile-phase conditions and pH to quickly screen multiple mobile-phase conditions

HSS Technology Advantages High-pressure-tolerant silica - Silica used for lower pressure HPLC separations is fragile at high pressure. HSS columns are the only commercially available

silica-based column that has been designed for UPLC. C and olumns Unique silica bonding - The silica substrate used for XSelect HSS columns combined with optimized ligand bonding gives the chromatographer a versatile and complimentary set of column choices. Whether your method requires 100% aqueous mobile phase or alternative retention mechanisms including reversed phase, normal phase, and HILIC, there is an XSelect HSS column tailored for performance.

Stationary Phase Characteristics onsumables

CSH CSH Ligand Type CSH C HSS C HSS C SB HSS T3 HSS PFP HSS CN 18 Fluoro-Phenyl Phenyl-Hexyl 18 18

Particle Size 2.5, 3.5, 5 µm 2.5, 3.5, 5 µm 2.5, 3.5, 5 µm 2.5, 3.5, 5 µm 2.5, 3.5, 5 µm 2.5, 3.5, 5 µm 2.5, 3.5, 5 µm 2.5, 3.5, 5 µm

Ligand Density* 2.3 µmol/m2 2.3 µmol/m2 2.3 µmol/m2 3.2 µmol/m2 1.6 µmol/m2 1.6 µmol/m2 3.2 µmol/m2 2.0 µmol/m2 Carbon Load* 15% 10% 14% 15% 8% 11% 7% 5% Endcap Style Proprietary None Proprietary Proprietary None Proprietary None None pH Range 1–11 1–8 1–11 1–8 2–8 2–8 2–8 2–8 Low pH 80 °C 60 °C 80 °C 45 °C 45 °C 45 °C 45 °C 45 °C Temp. Limit High pH 45 °C 45 °C 45 °C 45 °C 45 °C 45 °C 45 °C 45 °C Temp. Limit Pore Diameter* 130Å 130Å 130Å 100Å 100Å 100Å 100Å 100Å Surface Area* 185 m2/g 185 m2/g 185 m2/g 230 m2/g 230 m2/g 230 m2/g 230 m2/g 230 m2/g

*Expected or approximate value.

129 Enhanced Selectivity Reproducibilty Method development scientists use selectivity and retentivity to influence One of the challenges in designing stationary phases with large chromatographic behavior. Offering a family of chromatographic selectivity differences is maintaining acceptable batch-to-batch sorbents that provide diverse selectivities was one of the design reproducibility. Waters R&D recognized that method developers considerations for XSelect HPLC columns. need reproducible columns that will maintain performance over the lifetime of the method. This was a key consideration for the XSelect Waters continuing effort to develop LC particle technology leads to the column family. The figure below is an overlay of gradient separations innovations behind CSH and HSS particle development. No other family on columns containing nine different batches of XSelect CSH Fluoro- of LC products is focused on the development to maximize selectivity Phenyl materials. In these results, the batches include three different differences compared to commercially available chromatographic particle sizes demonstrating that even across different particle sizes, packing materials. The Selectivity Factor (S-value) measures how different maintain the same selectivity. The unique XSelect Fluoro-Phenyl (or similar) the selectivities of two column chemistries are under a given phase establishes an expectation that highly selective stationary set of conditions. The value is determined by measuring and plotting phases can be reproducible. the retention factors (k) of analytes of the two columns. Higher S-values indicate a higher degree of orthogonality.

2 XSelect CSH Fluoro-Phenyl 5 ìm Selectivity (S) = 100 x 1-R 8 10 2 6 1 5 4 7 9 Chemistries Engineered to Provide Wider Range of Selectivities 3

XSelect CSH C18 XSelect CSH Fluoro-Phenyl XSelect CSH Phenyl-Hexyl

10 XSelect CSH Fluoro-Phenyl 3.5 ìm ases onsumables H Ph CS ect el XS on

g k 5

R2 = 0.76 olumns and C S = 49

0 ACQUITY UPLC CSH Fluoro-Phenyl 1.7 ìm 05 10 15

kg on XBridge C18

XSelect HSS C18 XSelect HSS CN

XSelect HSS C18 SB XSelect CSH C18

XSelect HSS T3 XSelect CSH Fluoro-Phenyl

nalytical HPLC C XSelect HSS PFP XSelect CSH Phenyl-Hexyl 15 A

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 min

g) ACQUITY UPLC CSH Fluoro-Phenyl (1.7 µm) and XSelect CSH Fluoro-Phenyl (3.5 and 5 µm) (k 10 reproducibility and scalability. Gradient separations on 2.1 x 50 mm columns containing 9

ases different batches of CSH Technology Fluoro-Phenyl representing three (1.7, 3.5, and 5 µm) Ph particle sizes. Gradient: A: 15.4 mM ammonium formate, pH 3; B: acetonitrile; 5–90% B ect el linear in 5 minutes. Temperature: 30 °C. Injection volume: 5 µL. Detection: 254 nm. Flow XS rate: 0.5 mL/min. Analytes: (1) thiourea; (2) resorcinol; (3) metoprolol; (4) 3-nitrophenol; n on (5) 2-chlorobenzoic acid; (6) amitriptyline; (7) diethylphthalate; (8) fenoprofen; tio n 5 te (9) dipropylphthalate; (10) pyrenesulfonic acid. System: ACQUITY UPLC. Re

R2 = 0.63 S = 61

0 0510 15

Retention on BEH C18 (kg)

The Selectivity Factor (S-value) can be used to assess (and hence, help to create) orthogonality of selectivities between two different columns. Simply plot retention factors of one column versus another column, calculate the coefficient of determination (R2) and calculate the Selectivity Factor as indicated. Larger values indicate higher orthogonality.

130 www.waters.com/order Columns Designed for Method Development and Method Transferability Many laboratories function as teams that are a part of multi-national/multi-site organizations. From a global business perspective, it is vital to be able to quickly and easily develop robust methods that are not only compatible with all modern chromatographic detection modes, but are also transferable to laboratories and sites that may operate different LC system platforms. ACQUITY UPLC and XSelect columns, as part of the ACQUITY UPLC H-Class total system solution, were strategically created for the 21st-century global chromatographic marketplace. Using the same technology throughout the entire process, methods can be developed quickly with ACQUITY UPLC columns and then transferred to XSelect HPLC and preparative-scale XSelect OBD columns for isolation and purification.

Isolation and Purification

LC Conditions Preparative Conditions Injection Vol.: 1.25 mL

Columns: ACQUITY UPLC CSH C18, 2.1 x 50 mm, 1.7 μm Column: XSelect CSH C18 OBD Prep, Sample Conc.: 80 mg/mL for crude mix Part Number: 186005296 19 x 100 mm, 5 μm Diphenhydramine: 40 mg/mL Mobile Phase A: 0.1% formic acid in water Part Number: 186005421 Oxybutynin: 16 mg/mL Mobile Phase B: 0.1% formic acid in acetonitrile Mobile Phase A: 0.1% formic acid in water Terfenadine : 24 mg/mL Gradient: Time Flow Profile Mobile Phase B: 0.1% formic acid in acetonitrile Sample Diluent: DMSO (min) (mL/min) %A %B Curve Gradient: Time Flow Profile Curve Column Temp.: 40 °C Initial 0.9 95 5 Initial (min) (mL/min) %A %B Weak Needle Wash: 95/5 water/methanol 2.0 0.9 5 95 6 1.21 25 95 5 Initial Strong Needle Wash: 95/5 methanol/water 2.5 0.9 5 95 6 1.71 25 95 5 6 Detection: UV @ 220 nm 2.51 0.9 95 5 6 7.02 25 78.8 21.2 6 Sampling Rate: 1 point/sec 3.50 0.9 95 5 6 7.21 25 70.8 29.2 6 Filter Time Constant: 1 sec Injection Vol.: 1 μL 8.21 25 5 95 6 System: Waters 2525 Binary Gradient Module, Sample Conc.: 800 µg/mL for crude mix 8.31 25 95 5 6 2767 Sample Manager, Column Fluidics Sample Diluent: 50/50 methanol/water 13.21 25 95 5 6 Organizer, 2996 Photodiode Array Column Temp.: 40 °C Detector, ZQ™ Mass Spectrometer A

Weak Needle Wash: 95/5 water/acetonitrile C HPLC nalytical Strong Needle Wash: 95/5 acetonitrile/water 3. Run preparative separation using focused gradient Detection: UV @ 220 nm 1 3 Sampling Rate: 20 points/sec 2.00 Filter Time Constant: 0.1 sec Collected Fraction Compounds: System: ACQUITY UPLC with 1. Diphenhydramine 1.50 ACQUITY UPLC PDA 2. Oxybutynin

3. Terfenadine

AU 1.00

0.50 Prep Purification 2 100 mg total load C and olumns 2. Scale UPLC separation to preparative column scale 0.00 4. Collect and isolate fraction(s) of interest 23456 78 9 10 min

3 1.50 Target Analyte 1.50 1

3 1.00 1.00 AU AU Crude mix 1 Pooled purified fractions

0.50 Diphenhydramine = 56.3% purity 0.50 Diphenhydramine = 100% purity onsumables 2 2

0.00 0.00

0.0 0.20.4 0.6 0.81.0 1.21.4 1.6 1.8 2.0 min 0.0 0.2 0.40.6 0.81.0 1.21.4 1.6 1.8 2.0 min

1. Develop method using UPLC Technology 5. Confirm fraction purity using UPLC Technology

Using CSH Technology throughout the entire process, methods can be developed quickly with ACQUITY UPLC CSH columns and UPLC Technology and then transferred to preparative-scale XSelect OBD columns for isolation and purification. The purity of the isolated fraction(s) can then be measured/confirmed using ACQUITY UPLC CSH columns and UPLC Technology.

High mass loading applications like compound purification and dissolution testing are highly demanding of column performance. Under these extreme loading conditions, the limiting factor for reproducible scaling, often results from the inability to maintain peak shape. The combination of Optimum Bed Density (OBD) preparative columns with the benefits of XSelect particle technology gives the greatest benefit to the purification scientist. CSH Technology, for example, offers the highest loading of basic compounds using highly volatile, low-pH mobile phases often preferred by isolation and purification scientists. This increased loadability allows the use of narrower, lower volume, preparative columns thereby reducing solvent consumption and fraction volumes. Did you know... TruView LCMS Certified vials minimize analyte adsorption and are ideal for working with low sample concentrations. For more information, see page 68.

131 XBridge Columns With industry leading high pH stability and the highest level of chromatographic performance, XBridge BEH columns are the benchmark for LC method development. With a selection of 19 general purpose and application-specific stationary phases in the widest range of particle sizes, no other LC column family can match the versatility of the XBridge suite. Based on proven hybrid technology, the patented BEH particle* is a product of Waters commitment to provide you with the tools you need to solve the most demanding of chromatographic challenges. Leading separations scientists need LC columns that provide the highest level

of chromatographic performance to: Maximize efficiency  Provide long column lifetimes with increased high pH stability See page 170 Improve column reliability for part numbers and *US Patent # 6,686,035 B2 ordering information.

Stationary Phase Characteristics

Monofunctional BEH130 C18 BEH300 C18 BEH300 C4 BEH C18 Trifunctional Trifunctional Trifunctional Ligand Type Embedded HILIC Amide (Peptide (Peptide (Protein (Oligonucleotide C18 C8 Polar Group C6 Phenyl Separation Separation Separation Separation (Shield RP18) Technology) Technology) Technology) Technology) 2.5, 3.5, 2.5, 3.5, 2.5, 3.5, 2.5, 2.5, 2.5, 3.5, 5, 3.5, 5, onsumables Particle Size 3.5 µm 2.5 µm 5, 10 µm 5, 10 µm 5, 10 µm 3.5, 5 µm 3.5, 5 µm 3.5 µm 10 µm 10 µm Ligand Density** 3.1 µmol/m2 3.2 µmol/m2 3.3 µmol/m2 3.0 µmol/m2 N /A 7.5 µmol/m2 3.1 µmol/m2 3.1 µmol/m2 2.4 µmol/m2 3.1 µmol/m2 Carbon Load** 18% 13% 17% 15% unbonded 12% 18% 12% 8% 18% Endcap Style Proprietary Proprietary TMS Proprietary n/a None Proprietary Proprietary None Proprietary pH Range 1–12 1–12 2–11 1–12 1–9 2–11 1–12 1–12 1–10 1–12 olumns and C Low pH 80 °C 60 °C 50 °C 80 °C 60 °C 90 °C 80 °C 80 °C 80 °C 80 °C Temp. Limit High pH 60 °C 60 °C 45 °C 60 °C 45 °C 90 °C 60 °C 60 °C 50 °C 60 °C Temp. Limit Pore Diameter** 130Å 130Å 130Å 130Å 130Å 130Å 130Å 300Å 300Å 130Å Surface Area** 185 m2/g 185 m2/g 185 m2/g 185 m2/g 185 m2/g 185 m2/g 185 m2/g 90 m2/g 90 m2/g 185 m2/g ** Expected or approximate value. nalytical HPLC C A Maximizing Column Efficiency

One of the most important parameters in the design of the new hybrid van Deemter Curve Comparison particle was to significantly improve the chromatographic equivalence 12 to state-of-the-art silica based materials. The origins of band spreading, Reduced van Deemter Plot Terms which decreases separation efficiency, are described by the van Deemter a b c r2 10 XBridge C18 5 µm 1.04 8.7 0.044 0.998 equation: Symmetry C18 5 µm 0.76 10.6 0.045 0.999 Ht 8 SunFire C18 5 µm 0.95 11.6 0.040 0.997 1/ 2 N

ing Compound: Mobile Phase: 70/30 ACN/water us

h 6 Decanophenone Temperature: 30 °C h = a + b/u + cu

2 Data for both silica and BEH Technology packings, fitted to the 020406080

van Deemter equation demonstrate effective equivalence in u[uidp/Dm] efficiency. The c term for XBridge C18 is virtually identical to that The reduced plate height, h, is a function of the reduced linear velocity, u, [both normalized for the two state-of-the-art C18 silica columns, indicating that all for particle size] and a, b, and c summarize the contributions of eddy diffusion, longitudinal diffusion, and the sum of stationary- and mobile-phase mass transfer terms, respectively. these columns are comparable in mass transfer characteristics.

132 www.waters.com/order pH Stability

Improved Low pH Performance High pH Endurance The major cause of poor column lifetime in low pH mobile phases XBridge columns are engineered to be the most pH stable high-performance is due to the acid hydrolysis of the bonded phase. XBridge packings chromatographic phases commercially available. Approaches that claim incorporate state-of-the-art, proprietary procedures for bonding high pH resistance due to special surface modifications, cannot match the and end capping resulting in ligand stability and chromatographic lifetime of an XBridge column. Stability is the product of the BEH particle reproducibility at low pH. Compared to conventional materials, using synthetic process. an accelerated low pH stability test, XBridge C columns show very 18 Under accelerated pH 10 stability test conditions, a direct comparison little retention loss and exhibit exceptional column lifetime without to some of the most popular chromatographic phases, claimed to have resorting to sterically hindering the stationary phase. extended high pH stability, clearly shows the XBridge C18 column Accelerated Low pH Stability Tests of Competitive Columns lifetime exceeding that of the best columns by over 1000% with very little degradation in chromatographic performance. Probe: Methylparaben 100 XBridge C18 Accelerated High pH Stability Test of Competitive Columns

® Zorbax SB-C18 Compound: Acenaphthene 90

XBridge Phenyl 110

n 80 io Zorbax SB Phenyl nt A te 90 XBridge C18 HPLC C HPLC nalytical l Re ®

ia Luna C18(2) it 70 5s XTerra MS C18 N

% In ® ® Gemini™ C ACT Ace C18 70 18 % Initial ® 60 Luna C18 (2) ™ Gemini C18 ® ® YMC-Pak Pro C18 50 ® ® Capcell PAK MG II Zorbax Extend C18 50 050100 Hours in 1% TFA in H O pH 1.0 at 80 °C 2 30 0 50 100 150 200 hours

Comparative separations may not be representative of all applications. C and olumns Hours in 50 mM TEA, pH 10, 50 °C

Compounds: 1. Uracil 2. Propranolol 3. Naphthalene 4. Acenaphthene 5. Amitriptyline

1 3 Column Lifetime and Phosphate Buffer 2 4 5 Phosphate buffers are well recognized and are the most commonly N=13600 Initial accepted mobile-phase additive. Above pH 7, phosphate buffers are N=13500 100 h very aggressive and even the best reversed-phase silica columns onsumables are unable to withstand the alkaline conditions. The combination of N=14100 200 h phosphate, intermediate pH, and relatively low temperatures (40 °C) is well recognized to lead to short column lifetimes for most modern N=11700 300 h phases. XBridge BEH columns overcome this limitation and provide you with extended pH range and control over your separation. 04812 16 20 24 min Chromatograms, run at regular intervals during the high-pH lifetime study, verify Chromatographic Lifetime Comparisons in Phosphate Buffer that 86% of the original XBridge column efficiency remains after 300 hours at pH 10 and elevated temperature, with little change in peak shape or retention time.

® ™ Waters XBridge C18 Phenomenex Gemini C18 Comparative separations may not be representative of all applications. 300 Hours 30 Hours

N = 11700 N = 2218

024 8 12 16 0 24 min 0210 0 30 min 50 mM TEA, 50 °C, pH 10

Waters XBridge C18 Phenomenex Gemini C18

270 Hours 13 Hours

015 10 5 20 min 015 10 5 20 min

30 mM KH2PO4, 30 °C, pH 12.3 Aggressive alkaline mobile phases can quickly dissolve particle substrates creating voids in the packed bed. The BEH particle is chemically resistant allowing XBridge HPLC columns to perform under extreme mobile-phase conditions.

Comparative separations may not be representative of all applications.

133 Method Development Flexibility Selecting the most suitable column and separation conditions can be Changing mobile-phase pH is the single most effective strategy to extremely difficult. The XBridge family of columns is designed to eliminate influence analyte selectivity and retention. XBridge columns allow you the compromises of sorbent selection and deliver the flexibility to to use the full range of pH to control the elution of your target analytes. work under any mobile phase, temperature and pH conditions, thus speeding up the process to an optimized and rugged final method. Creating Selectivity Changes with Phosphate Buffer

A simple reversed-phase method development approach consists of Column: XBridge C18, 4.6 x 100 mm, 3.5 µm three XBridge columns, two mobile-phase pHs, and two organic Part Number: 186003033 Mobile Phase: A1: 30 mM potassium phosphate buffer (pH 2) solvents, that quickly defines the starting conditions for the most A2: 30 mM potassium phosphate buffer (pH 7) challenging of sample mixtures. An optimized approach to method A3: 30 mM potassium phosphate buffer (pH 12) Mobile Phase B: Acetonitrile development that quickly resolves complex mixtures saves you time, Flow Rate: 1.4 mL/min Gradient: Time Profile effort, and expense. (min) %A %B 0.0 90 10 7.0 20 80 8.0 20 80 XBridge Low pH in Methanol XBridge High pH in Methanol Injection Vol.: 20 µL Compounds: Sample Conc.: 50 µg/mL each in 1. Doxylamine 80/20 water/methanol 10 11 11 C18 C18 2. Benzamide 16.68 Column Temp.: 30 °C 6 10 8,13 2 9 2 3. Hydroxyisophthalic Acid 5 Detection: UV @ 210 nm (pH 2,7); 3 12 7 4. Doxepine 3 6 9 220 nm (pH 12) 8 5. Flavone 5 System: Alliance 2695 with 2996 PDA 1 4 7 1 6. Fenoprofen

10 11 11 Shield RP18 12 Shield RP18 6 13 1,2 8,9 2 9,10 5 7 8 3 12 3 4 5 3 6 2 pH 2 6 5 4 7 1

11 1

Phenyl 10 11 Phenyl 6 13 2 2 5 7,8 9 7 12 3 12 3 onsumables 6 9 10 5 1 4 1

048 12 16 20 min 048 12 16 20 min 4 3 6 5 2 pH 7 1 XBridge Low pH in Acetonitrile XBridge High pH in Acetonitrile

olumns and C 11 C18 C18 9,10 11 6 2 2 5 13 12 9 8 4 3 12 pH 12 3 7 3 2 6 5 10 1 6 1 4 7 5 8 4

Shield RP18 10 11 Shield RP18 6 1 2 2 13 5 12 9 3 8 3 12 7 6 4 9 10 1 1 4 7 5 8

11,12 0 12345678 min nalytical HPLC C Phenyl Phenyl 11 9 6 4,5 2 A 2 13 8 4 9 3 7 3 12 XBridge columns are unique in their ability to withstand these aggressive conditions 6 7 10 1 10 1 58 (even at pH 12) allowing complete flexibility for the method development chemist, while

048 12 16 20 min 048 12 16 20 min maintaining the highest efficiency values associated with silica columns.

Did you know... Waters now offers Suitability Standards to help benchmark and trend analytical system performance. For more information, see page 310.

134 www.waters.com/order XBridge HILIC Columns for Orthogonal Selectivity  HILIC techniques are gaining popularity as part of HPLC method development strategies. As a See page 170 complement to traditional reversed-phased chromatography, XBridge HILIC columns separations not only for part numbers and retain highly polar compounds but can provide large selectivity differences where it is possible to have ordering information. complete reversal of elution order compared to a reversed-phase separation.

3 HO 2 O H

NCH3 1 O

O Reversed Phase 1. 6-acetyl morphine V0 =0.25 XBridge C18

O H NCH 3 3 HO

2. morphine 1 2 HO O O V0 =0.26 OH HILIC O A XBridge HILIC HO HO O H

NCH3 C HPLC nalytical HO 3. morphine 3-ß-D-glucuronide 01 23 45 6 min

Due to the multi-modal retention mechanisms in HILIC, an orthogonal selectivity is observed compared to reversed phase, where the glucuronide metabolite of morphine elutes after morphine

under HILIC conditions. XBridge Amide Columns for Sugar Analysis  C and olumns XBridge Amide columns utilize a chemically stable, trifunctionally-bonded amide phase, enabling a new See page 170 dimension in stability and versatility for HILIC separations. In addition to enhanced retention of highly for part numbers and polar compounds, these columns are equally suited for sugar analysis. Some of the benefits are: ordering information. Enables high resolution, high speed analysis of carbohydrates in complex sample matrices while maintaining or improving chromatographic resolution.

Increased chemical stability enables the use of high pH and high temperature to collapse anomers without the loss of reducing sugars. onsumables BEH particle technology, in combination with a trifunctionally-bonded amide phase, provides exceptional column lifetime, thus improving assay robustness. Unlike amine-based columns used for carbohydrate analysis, the XBridge Amide column is not susceptible to Schiff-base formation, thus improving quantitation accuracy.

XBridge Amide Analysis of Saccharides

3 Column: XBridge BEH Amide, 4.6 x 150 mm, 3.5 µm Compounds: Part Number: 186004869 1. Fructose 10 µg/mL Mobile Phase A: 30/70 acetonitrile/water with 0.10% ammonium hydroxide 2. Glucose 10 µg/mL Mobile Phase B: 80/20 acetonitrile/water with 0.10% ammonium hydroxide 3. Sucrose 10 µg/mL Flow Rate: 0.4 mL/min 4. Maltose 10 µg/mL Gradient: Time Profile 5. Maltotriose 10 µg/mL (min) %A %B 6. Maltotetraose 10 µg/mL Initial 0 100 7. Maltopentaose 10 µg/mL 4 30.6 60 40 8. Maltohexahose 10 µg/mL 30.7 0 100 6 9. Maltoheptaose 10 µg/mL Inj. Vol.: 10 µL 2 Sample Diluent: 50/50 acetonitrile/water 7 Column Temp.: 35 °C 1 System: ACQUITY UPLC with ACQUITY UPLC TQD SIR m/z: 179.2 (fructose, glucose); 341.3 (sucrose, maltose); 503.4, 8 665.5, 827.6, 989.7, 115 (maltooligosaccharides [n=1 to 5]) 5

042 6810 12 14 16 18 20 22 24 26 28 30 32 min

135 SunFire Columns SunFire columns set the standard for the state-of-the-art bonded

C18 and C8 silica HPLC columns. Benefiting from years of research and product development, these columns represent the best in particle and bonding expertise and deliver an industry-leading level of chromatographic performance.

Stationary Phase Characteristics

Particle Pore Carbon Chemistry Particle Size End Capped Shape Size Load

C18 2.5, 3.5, 5, 10 µm Spherical 100Å 16% Ye s

C8 2.5, 3.5, 5, 10 µm Spherical 100Å 12% Ye s

Silica 5, 10 µm Spherical 100Å Unbonded n/a 

Excellent Low pH Stability See page 164

100 for part numbers and ordering information. 90

SunFire C18 80

70 GL Sciences® Inertsil® ODS-3 ® ® ACT Ace C18 % of Initial Retention Time 60 onsumables ® ® Phenomenex Luna C18 (2) 50 020406080

Hours in 1% TFA in H2O at 80 ˚C Stability at low pH improves SunFire column lifetimes compared to those of other silica-based HPLC column brands. olumns and C

Superior Peak Shape and Efficiency With the bonding and end capping technologies for the SunFire columns, Waters developed a sorbent with superior peak shape performance. SunFire columns provide symmetrical peaks for improved resolution and quantitation of acidic, neutral, and basic compounds at low- and intermediate-pH ranges.

nalytical HPLC C Comparison of C 5 µm HPLC Columns Isocratic Separation Compounds: 18 A Mobile Phase A: 35% 20 mM dipotassium phosphate/ 1. Uracil 20 mM monopotassium phosphate pH 7.0 2. Propranolol Mobile Phase B: 65% methanol 3. Butylparaben SunFire C18 ® ® Wavelength: 254 nm 4. Naphthalene Phenomenex Luna C18 (2)

Flow Rate: 1.0 mL/min 5. Acenaphthene ® ™ Injection Vol.: 14 µL 6. Amitriptyline Thermo Scientific HyPURITY Elite C18 ® ® Column Temp.: 23 ˚C ACT Ace C18 System: Alliance 2695 with 2487 Dual GL Sciences® Inertsil® ODS-3 Wavelength Absorbance detector 2 ® 1 YMC-Pro C18

3 Agilent® Purosphere® RP-18E

® ® Agilent Zorbax Eclipse XDB C18 4 SunFire C18 TUSP – 1.26 5 6 4.6 x 150 mm, 5 µm 1.0 1.5 2.0 2.5 3.0 3.5 4.0 Part Number: 186002559 USP Tailing Factor (TUSP)

® ® In this comparison test of columns packed with 5 µm particles, the SunFire C18 column has TUSP – 1.35 Phenomenex Luna C18 (2) 4.6 x 150 mm, 5 µm the lowest USP tailing factor for amitriptyline using pH 7 mobile-phase conditions. Comparative separations may not be representative of all applications.

® ® TUSP – 1.79 ACT Ace C18 4.6 x 150 mm, 5 µm

0 5 10 15 20 25 30 35 40 min

Comparative separations may not be representative of all applications.

136 www.waters.com/order Batch-to-Batch Reproducibility In establishing new analytical methods for the latest pharmaceutical and biopharmaceutical products, the selection of a reproducible HPLC column is essential. The selected column needs to provide the same chromatographic results over the life of the method and the new drug product. SunFire columns have demonstrated superior reproducibility over many years. Batches randomly selected over a four-year period show excellent reproducibility in the example below.

Batch 112 (2005)

Batch 118 (2007)

Batch 130 (2007)

Batch 134 (2008) A HPLC C HPLC nalytical

0510 15 20 25 30 35 40 min

This excellent reproducibility is a result of our commitment to maintaining the tightest specifications in the HPLC column industry. SunFire columns start with high purity raw materials and are produced using tightly controlled manufacturing processes and column packing procedures that provide today’s scientists with the best, most reproducible HPLC columns available.

Did you know... Did you know... C and olumns The recommended column cleaning and regenerating Waters Suitability Standards can help you benchmark procedures are listed in the Care and Use Instructions. your system performance and help in trending results. For brand-specific care and use instructions, visit For more information, see page 310. www.waters.com/chemcu onsumables

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137 Atlantis Columns The Atlantis family of HPLC columns was created to solve one of the most challenging chromatographic problems, retaining polar compounds. Through historical experience and synthetic innovation, Atlantis columns lead the industry in providing exceptional performance, versatility, retention and chromatographic stability for polar compounds, while also affording balanced retention for broad analyte mixtures. For retention and separation of polar compounds via reversed-phase HPLC, Atlantis T3 columns set the standard for polar compound retention. Atlantis T3 columns are a universal,

silica-based, reversed-phase C18 line of HPLC columns that not only retain and separate small, water-soluble polar organic compounds, but also provide superior performance across a wider pH range. Atlantis columns are the culmination of over 30 years of bonded

phase research and are a vast improvement upon the highly successful Atlantis dC18 line of columns. For reversed-phase applications, Atlantis T3 columns should be considered the first choice when developing a separation of polar (and non-polar) compounds. The advantages

of T3 bonding technology is extended to UPLC in the ACQUITY UPLC HSS T3 columns. Stationary Phase Characteristics  Intended Use HPLC See page 161 Particle Type Atlantis Silica

Available Chemistries T3, dC18, HILIC Silica for part numbers and

pH Range T3: 2–8; dC18: 3–7; HILIC Silica: 1–5 ordering information. Maximum Rated Pressure 6000 psi [∼400 bar] Particle Size 3, 5, 10 �m Pore Diameter / Volume 100Å/1.0 mL/g Surface Area 330 m2/g onsumables Superior Low pH Stability Long Column Lifetimes at Low pH

Atlantis T3 The creation of highly retentive, reversed-phase 20 hr Exposure to 0.5% TFA at 60 ˚C C18 columns for polar compound retention involves SunFire C18 bonding at a ligand density that is less hydrophobic Phenomenex® Synergi™ Hydro-RP and, therefore, more compatible with the weak, highly ® ® ® Shiseido Capcell Pak C18 AQ olumns and C aqueous mobile phases required for retaining polar Phenomenex Synergi Polar-RP compounds. When exposed to strongly acidic mobile Agilent® Zorbax® Eclipse® Plus phases (i.e. < pH 2.0), traditional C18 columns can Atlantis dC 18 exhibit bonded phase hydrolysis resulting in gradual Phenomenex Aqua® loss of retention, loss of efficiency as well as change GL Sciences® Inertsil® ODS-SP in selectivity. 0102030405060 70% % Loss of Retention for Methyl Paraben Atlantis T3 columns resist ligand cleavage by

nalytical HPLC C utilizing a trifunctional attachment of the C18 T3 bonding allows Atlantis T3 columns to provide long column lifetimes under harsh, low pH conditions. A Comparative separations may not be representative of all applications. phase to the particle surface, thus providing exceptional column life at low pH.

Improved Peak Shape for Bases at pH 7 Improved pH 7 Performance At pH 7, poor peak shape for amine-containing Atlantis T3 bases and shortened column lifetimes are encountered GL Sciences® Inertsil® ODS-SP when using intermediate ligand density C18 ® ® ® Shiseido Capcell PAK C18 AQ columns designed for polar compound retention.

Phenomenex Synergi Polar-RP Poor peak shape is due to secondary interactions ® ™ Phenomenex Synergi Hydro-RP with unreacted silanols that remain present after ® Phenomenex Aqua bonding and endcapping. The proprietary T3 Atlantis dC18 endcapping procedure reacts with more of these ® ® Agilent Zorbax SB-Aq active silanols thereby dramatically improving ® ® Thermo Aquasil C18 Did not elute peak shape for bases. Shortened column lifetimes 1.0 1.5 2.0 2.53.0 3.54.0 4.55.0 are due to the dissolution of the silica particle Amitriptyline USP Tailing Factor substrate by the high-pH mobile phase. The more When compared to other reversed-phase ‘polar compound retention columns’, Atlantis T3 columns provide superior effective and efficient T3 bonding helps ‘protect’ pH 7 peak shape. the silica substrate from dissolution, thus providing Comparative separations may not be representative of all applications. longer column lifetimes.

138 www.waters.com/order Atlantis HILIC Silica Retention Mechanisms in Hydrophilic-Interaction Chromatography (HILIC) HILIC is an alternative chromatographic technique that offers complementary selectivity to RP and often retains very polar species

H2O ACN/H2O that cannot be retained by traditional means. Atlantis HILIC Silica (90:10) 90% ACN H H 90% ACN H N Si N columns were designed to retain very polar, organic molecules that O N O H O- Si O O N Si N O 1 Si O- O Si are too polar to retain by RP. Unlike the highly aqueous mobile N O N H + O O O 3 N N Si N H + H O H Si Si 3 Si + O O NH O H-O phases required for polar retention in RP separations, Atlantis HILIC + 3 NH Si H Si 3 O-H O O- O O O- H H O Si Silica columns employ highly volatile (>80% organic) mobile phases H-O Si O Si O Si H H O-H O O H O H H O- O which are ideal for mass spectrometry (MS) response and sensitivity. Si O Si Si O O H Si O O- H O O-H H-O O Si 2 H Additionally, direct compatibility with high organic SPE eluates Si Si O- Si O O O O O Si O Si Si H O O dramatically increases the number of samples that can be handled, Water Si Si Si O O H O Si O O H Si H O Si O O-H O O- Si Si O Si thus substantially increasing sample throughput. Acetonitrile/Water (90:10) Si O O- O-H O O Si Si Si Si Si O O O O O O O Si HILIC retention mechanisms are a complex combination of partitioning, Si Si Si ion-exchange and hydrogen bonding, resulting in enhanced retention for polar analytes.1,2 Atlantis HILIC Silica columns, used in combination with high organic (>80% acetonitrile) mobile phases, result in retention The combination of partitioning and weak cation exchange results in retention of polar of analytes that are simply too polar to retain by traditional RP bases with Atlantis HILIC Silica columns. chromatography.

E.S. Grumbach et al LCGC N. Am. 2004, 22, 1010-1023 A 2 E.S. Grumbach et al J. Sep. Sci. 2008, 31, 1511-1518 HPLC C HPLC nalytical

Guide to Retaining Polar Compounds

Is the polar compound an acid, neutral C and olumns Base or base? Acid or Neutral

XSelect HSS T3, Ye s Can it be Can it be Ye s Atlantis T3 Atlantis T3 retained by retained by or or reversed-phase? reversed-phase? XSelect HSS T3 XBridge C18 No No Ye s onsumables

Sufficient mass Atlantis HILIC Silica, Insufficient spectrometry XBridge HILIC retention sensitivity? or XBridge Amide

Ye s

No Alternate separation technique, such as Key: ion-pair or Atlantis T3 = low pH (pH 3), high aqueous (>95%) ion-exchange

XBridge C18 = high pH (pH 10), high aqueous (>95%) Atlantis HILIC Silica = low pH (pH 3), 10 mM ammonium formate with 0.2% formic acid in 95% acetonitrile/5% water XBridge HILIC = mid pH (pH 5), 10 mM ammonium acetate with 0.02% acetic acid in 95% acetonitrile/5% water

XBridge Amide = mid pH (pH 5), 10 mM ammonium acetate with 0.02% acetic acid in 95% acetonitrile/5% water

XSelect HSS T3 = low pH (pH 3), high aqueous (>95%) 

See page 161 for part numbers and ordering information.

139 XTerra Columns Silica-based packing materials limit the chromatographic potential for separation scientists. Restrictions on speed, resolution, pH, temperature, and loading capacity are imposed upon the chromatographer by limitations of the stationary phase material. The patented* Hybrid Particle Technology of XTerra columns allows chromatographers to break these boundaries and realize the full potential of their analytical and preparative separations. * Patent # 6,686,035 B2

Stationary Phase Characteristics Particle Pore Carbon End-  Packing Particle Size(s) Shape Size Load capped See page 174 XTerra MS C18 Spherical 2.5, 3.5, 5, 10 µm 125Å 15.5% Ye s for part numbers and XTerra MS C8 Spherical 2.5, 3.5, 5, 10 µm 125Å 12.0% Ye s ordering information. XTerra RP18 Spherical 3.5, 5, 10 µm 125Å 15.5% Ye s XTerra RP8 Spherical 3.5, 5, 10 µm 125Å 13.5% Ye s XTerra Phenyl Spherical 3.5, 5 µm 125Å 12.0% Ye s

Hybrid Particle Technology Effect of XTerra Column Chemistry on Selectivity and Retention In Hybrid Particle Technology, one out of every three silanols is replaced onsumables Column: 4.6 x 150 mm, 5 µm Flow Rate: 1.0 mL/min with a methyl group during synthesis. This hydrophobicity is distributed Mobile Phase A: Water Injection Vol.: 15 mL Mobile Phase B: Methanol Column Temp.: 30 °C throughout the entire structure of the particle backbone. The result is Mobile Phase C: 50 mM formic acid, pH 2.45 Detection: UV @ 254 nm a rugged hybrid (inorganic/organic) particle that can be operated at Gradient: Time Profile System: Alliance 2695 (min) %A %B %C with 996 PDA high speeds, high temperatures, and high pH. The presence of 33% 0.0 40 50 10 fewer residual silanols (after endcapping and bonding) also means 20.0 25 65 10 Compounds: 35.0 25 65 10 1. Suprofen that XTerra columns give exceptionally sharp, high-efficiency peaks 2. Tolmetin

olumns and C 3. Naproxen for basic compounds. 3 4. Fenoprofen

XTerra MS C8 1 5. Ibuprofen 6 6. Diclofenac Part Number: 186000490 2

O 4 O Si O 5 Did you know... Guard columns can help extend the life 3

nalytical HPLC C XTerra RP8 of your analytical column. 1 6 A Part Number: 186000493 2

CH 4 See page 147 for product information. 3 OSi

CH 3 5

XTerra Phenyl 1 Part Number: 186001146 2 6

O 4

O Si CH2 CH 5 CH3 CH3

5101520 25 min The column stationary phase chemistries of XTerra columns are key components in an overall method development plan.

140 www.waters.com/order Symmetry Columns As chemists develop analytical methods for pharmaceutical and biopharmaceutical products, the selection of a reproducible HPLC column is essential. The selected column needs to provide the same chromatographic results over the life of the new drug product. Symmetry columns demonstrate superior reproducibility over many years. Reproducibility is our number one priority in supplying Symmetry columns. This excellent reproducibility is a result of our commitment to maintaining the tightest specifications in the HPLC column industry. Symmetry columns are engineered with high purity raw materials, tightly controlled manufacturing processes and column packing procedures that

provide scientists with the best, most reproducible HPLC column available. Our goal is to supply a family of HPLC columns that you can rely on for rugged and  robust methods. Symmetry columns let you increase your laboratory’s productivity, and See page 166 allow easier method transfer between labs and around the globe. for part numbers and ordering information.

Stationary Phase Characteristics

Packing Chemistry Particle Size(s) Particle Shape Pore Size Carbon Load End Capped A

C18 3.5, 5, 7 μm Spherical 100Å 19% Ye s C HPLC nalytical Symmetry C8 3.5, 5, 7 μm Spherical 100Å 12% Ye s RP18 3.5, 5, 7 µm Spherical 100Å 17% Ye s SymmetryShield RP8 3.5, 5, 7 µm Spherical 100Å 15% Ye s

The Benefit of Narrow Specification Ranges C18 Ligand Surface Coverage Specification Ranges

Symmetry columns have the narrowest ligand surface coverage C and olumns specifications in the industry. Surface coverage (ligand density) is 3.09 3.31 one of the most important parameters affecting reproducibility. The Symmetry C18 impact of a small variation in surface coverage on the amitriptyline/ 2.75 3.75 acenaphthene alpha specification for Symmetry C18 columns is shown Column X here. Narrow column specifications benefit the chromatographer in 2.6 no upper limit achieving reproducible results. The surface coverage requirements of Column Y onsumables Symmetry are the tightest specifications in the industry, resulting in minimal shifts in resolution and more reproducible batch-to-batch 1.5 2.0 2.5 3.0 3.5 4.0 and column-to-column results. 2 C18 Surface Coverage (µmol/m )

Narrow Specification Ranges Provide Smaller Shifts in Alpha (α) Minimal Changes in Resolution Means More

2.0 Reproducible Results*

1.8 4.14 2.86 1.6 Symmetry C18

1.4

1.2

Acenaphthene Specification Ranges

for Symmetry C18

Alpha for Amitriptyline/ 1.0

Specification X 0.8 4.70 0.81 1.5 2.0 2.5 3.0 3.5 4.0

2 C18 Surface Coverage (µmol/m )

Smaller Shifts in Alpha (α) Reduce Changes in Peak Resolution Specification Y 5.37 0

R = resolution √N α-1 K2 N = number of peaks R = ( ) ( ) ( ) α = alpha value Maximum Resolution Minimum Resolution 4 α K2+1 K = peak retention * Based on vendor’s column product specifications for ligand density

141 Long Lifetime Outstanding Packed Bed Stability—10,000 Injections

Columns that last longer save you money. Symmetry columns can last Column: Symmetry C8, 3.9 mm x 150 mm, 5 µm, over 10,000 injections with minimal loss in efficiency, minimal with Sentry Guard Column, 3.9 mm x 20 mm, 5 µm Part Numbers: WAT046970 (column) & WAT054250 increase in backpressure, and minimal change in retention time. (guard column) Mobile Phase: Water/methanol/glacial These columns deliver guaranteed consistent performance optimizing 1 acetic acid 79:20:1 3 Flow Rate: 1.0 mL/minute the two key factors that control column lifetime which are hydrolytic Injection Volume: 10 µL stability of the packing material and mechanical stability of the Temperature: 25 °C packed bed. Detection: UV @ 254 4 5 2 Compounds: Comprehensive Certificate of Analysis 6 1. Sulfanilamide 2. Sulfadiazine 3. Sulfathiazole 4. Sulfamerazine 5. Sulfamethazine 6. Succinylsulfathiazole

Injection 10,000

Injection 1 A three-page certificate of analysis is included with every Symmetry 0246810 min column. We report both our specifications, which are the tightest in The long-term mechanical stability of Symmetry columns is demonstrated in this analysis the industry, and the results of the 22 critical tests each column must of sulfa drugs. Over 10,000 injections were run with minimal loss in column efficiency. The use of the Sentry guard column is critical to achieving these types of column lifetimes. pass before it carries the Symmetry brand. onsumables SymmetryShield Columns Excellent Peak Shape SymmetryShield HPLC columns are an excellent choice to achieve superior peak shapes for basic compounds. These reversed-phase columns are based on Waters patented embedded polar group technology that literally ‘shields’ the silica’s residual surface silanols from highly basic analytes. olumns and C Shield Technology Industry Leader for Peak Shape of Basic Compounds

O Polar SymmetryShield RP18 1.2 Si Group Brand L 1.5

Brand HP 1.66

CH3 CH3 Brand HE 1.75 Reduce interaction with silanols nalytical HPLC C Improve water-wettability Brand DO 1.78 A Brand DA 1.9 The embedded polar group, which is in close proximity to the silica Brand I 1.95 surface, further reduces the activity of the surface silanols in Brand P 2.39

SymmetryShield packing materials. Brand E 3.6 As a result, SymmetryShield columns: 01234 Deliver significantly improved peak shape and resolution over USP Tailing Factor a broad pH range.

Exhibit less retention of basic compounds than conventional C18 In this comparison test, SymmetryShield RP18 has the lowest USP tailing factor for the and C8 columns. basic probe amitriptyline under the most demanding pH 7 mobile-phase conditions.

In fact, SymmetryShield columns deliver USP Tailing Factors of less than 1.2 over a pH range between pH 2–8. By delivering lower  limits of detection and quantitation, SymmetryShield columns will enable you to develop new methods faster and improve the productivity See page 166 of your laboratory. for part numbers and ordering information.

142 www.waters.com/order Unique Selectivity Perfect for Use with Highly Aqueous Mobile Phases SymmetryShield columns exhibit less retention for basic compounds SymmetryShield columns with Shield Technology are the perfect

due to lower silanol activity than conventional C18 and C8 columns. choice for applications that require low organic, highly aqueous SymmetryShield columns are an excellent methods development tool. mobile-phase conditions. These reversed-phase particles are water wettable allowing better interaction between the particles and your In the example shown, a leading pharmaceutical company was faced sample. The result is stable retention in highly aqueous mobile phases with a challenging separation. It was only when a SymmetryShield with superior peak shape. column was substituted for a conventional C8 column that a much better separation was achieved without having to change mobile- phase conditions. SymmetryShield Columns–No Pore Dewetting in Highly Aqueous Mobile Phases SymmetryShield Columns Deliver Unique Selectivity Standard Silica C8 Columns—Pore Dewetting

Mobile Phase: Aqueous SymmetryShield RP18 Column: SymmetryShield RP18, 3.9 x 150 mm, 5 µm Column: Standard C8 Silica Column 0.1% Acetic Acid Symmetry C18, 3.9 x 150 mm, 5 µm 4.6 x 150 mm, 5 µm Part Numbers: 186000108 (SymmetryShield); 1 2 WAT046980 (Symmetry) 4 Mobile Phase: 65% methanol; 35% 20 mM monopotassium phosphate/ Amoxicillin Initial dipotassium phosphate at pH 7 Flow Rate: 1.0 mL/min 3 Detection: UV @ 254 nm Column Temp.: 23 °C After pump stopped and re-started A (Pore dewetting: ~ 100%) Compounds: C HPLC nalytical 5 USP Tailing Factor = 1.2 AU 1. Uracil 6 2. Propranolol 3. Butylparaben 7 02468 10 min 4. Dipropylphthalate 5. Naphthalene No retention for analyte 6. Amitriptyline 7. Acenaphthene Most HPLC columns lose their retention when running low or 0% organic mobile phases if the flow is stopped. When this happens the bonded particle pores become de-wetted. The result is termed ‘pore dewetting’ and you lose the rententivity of your analyte. 0510 15 20 25 30 min C and olumns

Symmetry C18 SymmetryShield RP8 Columns—No Pore Dewetting! 1 2 Mobile Phase: Aqueous Column: SymmetryShield RP8 3 0.1% Acetic Acid 4.6 x 150 mm, 5 µm Amoxicillin Part Number: WAT200662 Initial 4 onsumables

No change in After pump stopped and re-started AU USP Tailing Factor = 1.9 5 retention time (Pore dewetting: ~ 3%)

7 6 0 2 4 6 8 10 min

SymmetryShield columns prevent this from happening because the embedded polar group maintains the wetted pore surface, ensuring retention of your analyte and eliminating the 0510 15 20 25 30 min need for a lengthy re-wetting and column re-equilibration process.

Did you know... Atlantis T3 and XSelect HSS T3 columns provide superior polar compound retention while being compatible with 100% aqueous mobile phases.

143 Waters Spherisorb Columns One of the great advantages of choosing Waters Spherisorb columns is the range of choices available to you. Waters Spherisorb columns are produced in a wide range of sizes and bonded phases to meet your chromatographic needs. Silica is produced in 3, 5, and 10 µm particle

sizes. The high quality bonded phases give many different and unique

separation selectivities. 

See page 168 for part numbers and ordering information.

Stationary Phase Characteristics

Particle Surface Pore Pore Particle Carbon Ligand End- Product Description Shape Area Size Volume Size Load Coverage capped Phases

2 2 ODS2 (C18) Spherical 220 m /g 80Å 0.50 ml/g 3, 5, 10 µm 11.5% 2.98 µMol/m Ye s

2 2 ODS1 (C18) Spherical 220 m /g 80Å 0.50 ml/g 3, 5, 10 µm 6.2% 1.49 µMol/m No

2 2 ODSB (C18) Spherical 220 m /g 80Å 0.50 ml/g 5 µm 11.5% 2.98 µMol/m Ye s

2 2 C8 Spherical 220 m /g 80Å 0.50 ml/g 3, 5, 10 µm 5.75% 3.12 µMol/m Ye s

2 2 C6 Spherical 220 m /g 80Å 0.50 ml/g 3, 5, 10 µm 4.7% 3.36 µMol/m Ye s

2 2 C1 Spherical 220 m /g 80Å 0.50 ml/g 3, 5, 10 µm 2.15% 2.97 µMol/m No onsumables 2 2 NH2 (Amino) Spherical 220 m /g 80Å 0.50 ml/g 3, 5, 10 µm 1.9% 2.64 µMol/m No Phenyl Spherical 220 m2/g 80Å 0.50 ml/g 3, 5, 10 µm 2.5% 1.72 µMol/m2 No CN (Nitrile) Spherical 220 m2/g 80Å 0.50 ml/g 3, 5, 10 µm 3.1% 3.29 µMol/m2 No OD/CN Spherical 220 m2/g 80Å 0.50 ml/g 5 µm 5% 1.15 µMol/m2 Ye s W (Silica) Spherical 220 m2/g 80Å 0.50 ml/g 3, 5, 10 µm — — —

olumns and C Ion Exchangers SCX Spherical 220 m2/g 80Å 0.50 ml/g 5, 10 µm 4% — No SAX Spherical 220 m2/g 80Å 0.50 ml/g 5, 10 µm 4% — No

Waters Spherisorb Analytical Columns Waters Spherisorb analytical columns come with industry standard endfittings. The threaded female fitting requires a Parker-style compression nalytical HPLC C

A screw and ferrule to connect the HPLC tubing to the analytical column. If your instrument is set up for Waters columns, replace your ferrule on the connector tubing and set it for the Parker standard.

Waters Spherisorb Cartridge Columns Waters Spherisorb cartridge columns require reusable cartridge endfittings 2 3 (Part No. PSS614100). These endfittings should be ordered the first time a cartridge is purchased. The reusable endfittings are a Parker-

style standard endfitting. Coupler fittings are also available to allow 1 you to link a guard cartridge to your analytical cartridge. 4

1. 30 mm Stand Alone Guard/Column (endfittings not included) 2. Extended endfitting for use with 10 mm Integral Guard: PSS614108 3. 10 mm Integral Guard Column 4. Column Coupler: PSS614102

144 www.waters.com/order Nova-Pak Columns The 4 µm small particle size of Nova-Pak bonded phases offers high resolution as well as more efficient and faster chromatography. T his is accomplished by using shorter columns, thereby reducing solvent consumption or by using longer columns to resolve even the most complex mixture. Analytical columns with 4 µm particle size packings

are available in 75, 150, and 300 mm length steel columns. Steel cartridge columns with reusable endfittings are available in 50,  100, 150, and 250 mm lengths. Semi-preparative columns with 6 µm See page 162 particle size packings give you an unparalleled range of separation for part numbers and possibilities. Stringent QC procedures in our cGMP manufacturing ordering information. facility ensure batch-to-batch reproducibility.

Stationary Phase Characteristics

Packing Chemistry Particle Size Particle Shape Pore Size Carbon Load End Capped

C18 4 µm Spherical 60Å 7% Ye s

C8 4 µm Spherical 60Å 4% Ye s Nova-Pak Phenyl 4 µm Spherical 60Å 5% Ye s A

CN HP 4 µm Spherical 60Å 2% Ye s C HPLC nalytical Silica 4 µm Spherical 60Å n/a n/a

C18 6 µm Spherical 60Å 7% Ye s Prep Nova-Pak HR Silica 6 µm Spherical 60Å n/a n/a

Nova-Pak Analytical Columns Prep Nova-Pak HR Columns C and olumns Analytical columns with 4 µm Nova-Pak packing material gives you Prep Nova-Pak HR 6 µm packing is available in both economical an unparalleled range of separation possibilities. column segments and preparative columns. The dimensions of the columns have been chosen specifically to make scale-up from analytical to preparative scales simple and fast. The high efficiency packing Nova-Pak Cartridge Columns gives you faster separations, using less solvent, with the added onsumables The cartridge column requires reusable finger-tightened endfittings advantage of more concentrated fractions, all of which reduce the which are purchased separately. When you change columns, just cost of your preparative chromatography. replace the cartridge and use the endfittings again. The cartridge column assembly includes one disposable cartridge column and reusable endfittings. Nova-Pak Analytical Method Validation Kit

The Method Validation Kit includes three Nova-Pak C18, 4.6 x 150 mm, 4 µm, steel columns. Two of these columns are packed with material from one particular batch of material and the other column is packed with material from a different batch of the bulk material. All three columns are identified with the batch number. The columns are ideal for testing the ruggedness of new applications for regulated methods. Nova-Pak batch-to-batch reproducibility is guaranteed.

145

Delta-Pak Columns µBondapak/Bondapak Packings

 Delta-Pak columns, ideal for the  If your method calls for separation of peptides, proteins, See page 160 µBondapak C18 packings, See page 160 and natural products, are based on for part numbers and there is only one µBondapak for part numbers and a highly stable, bonded, end- ordering information. C18 packing material. Almost ordering information. capped 5 µm or 15 µm spherical anything can be made once, but silica. Delta-Pak columns are making a packing material reproducible year-to-year since 1973 available in two different pore size materials demonstrates true manufacturing and QC capability. Many companies

(100Å and 300Å) with a C18 or C4 bonded phase. claim to have columns with ‘µBondapak-like selectivity’ but not one has ever passed Waters stringent QC batch test. Waters has set the Delta-Pak columns are available in different cartridge and column standard for reproducible, high-performance chromatographic columns. configurations, providing consistent and predictable scale-up capabilities Waters µBondapak columns are still the most widely referenced and for milligram-to-gram scale purifications. Also, by having the same requested columns in the world. chemistry available on a high-efficiency 5 µm particle, you can easily check the purity of your preparative-scale fractions. Stationary Phase Characteristics

Delta-Pak Columns Stationary Phase Physical Characteristics Particle Particle Pore Carbon End- Packing Chemistry Size Shape Size Load capped Particle Particle Pore Carbon End- Packing Chemistry Size Shape Size Load capped µBondapak C18 10 µm Irregular 125Å 10% Ye s µBondapak Phenyl 10 µm Irregular 125Å 8% Ye s C18 5 µm Spherical 100Å 17% Yes µBondapak CN 10 µm Irregular 125Å 6% Ye s C18 5 µm Spherical 300Å 7% Yes

µBondapak NH2 10 µm Irregular 125Å 3.5% No C4 5 µm Spherical 100Å 7% Yes

Bondapak C18 15–20 µm Irregular 125Å 10% Ye s C4 5 µm Spherical 300Å 3% Yes Delta-Pak C18 15 µm Spherical 100Å 17% Yes

onsumables C18 15 µm Spherical 300Å 7% Yes µPorasil/Porasil Silica Packings

C4 15 µm Spherical 100Å 7% Yes Waters µPorasil 10 µm  C4 15 µm Spherical 300Å 3% Yes packing material provides high efficiency and low backpressure. See page 160 Porasil silica packing material Purification of a Synthetic Peptide Using Delta-Pak C18 Packing for part numbers and is available in 15–20 and ordering information. Column: Delta-Pak C18, 5 µm, 300Å, 3.9 x 150 mm 37–55 µm particle sizes. olumns and C Part Number: WAT011793 Sample: Synthetic peptide-neurotensin (5 mg/mL) Stationary Phase Characteristics Injection: 10 µL (50 µg) Mobile Phase A: Water with 0.1% TFA Particle Particle Pore Carbon End- Mobile Phase B: Acetonitrile with 0.1% TFA Packing Chemistry Gradient: 0–2 min, 5% B Size Shape Size Load capped Conditions: 2–27 min, 5–9% B 30–31 min, 90–5% B µPorasil Silica 10 µm Irregular 125Å n/a n/a Flow Rate: 1 mL/min Porasil Silica 15–20 µm Irregular 125Å n/a n/a Detection: UV @ 230 nm nalytical HPLC C

A Resolve Columns  The non-endcapped Resolve packings are significantly different from Nova-Pak and Bondapak packing See page 162 materials. The change in chromatographic behavior is most commonly noticed with polar compounds for part numbers and which are typically more retained. Basic compounds can be chromatographed using mobile-phase ordering information.

modifiers such as ion-pairing reagents which reduce excessive tailing. Resolve C18 and Silica columns are available for applications requiring high resolution.

Stationary Phase Characteristics

Packing Chemistry Particle Size Particle Shape Pore Size Carbon Load End Capped Did you know... Silica 5 µm Spherical 90Å n/a n/a µBondapak has been

C18 5 µm Spherical 90Å 10% No manufactured to the same

C18 10 µm Spherical 90Å 10% No specifications since 1973! Resolve C8 10 µm Spherical 90Å 5% No CN 10 µm Spherical 90Å 3% No Silica 10 µm Spherical 90Å n/a n/a

146 www.waters.com/order Guard Products Waters Cartridge and Guard Column Guide Guide to Guard Column Selection and Use Guard Columns Guard columns are used primarily to protect expensive analytical and Universal Sentry Guard Holder (Use on any column) preparative columns by removing particulate and strongly retained sample components which ultimately decrease the lifetime of the analytical column. Although economy alone is a persuasive argument for the use of guard columns, the need for a long, stable life in the Universal Sentry Guard Holder analytical column in order to obtain reliable and reproducible results is perhaps even more important. Cartridge Columns Guard columns are installed between the injector and the analytical column of a liquid chromatographic system. Guard columns may also Cartridge End-Fittings be used to perform automated, on-line sample cleanup. In situations where a guard column is not needed, or where no suitable guard Cartridge with End-Fittings column is available, an in-line filter should be used to protect the analytical column from microparticulate contamination. The in-line filter is placed between the injector and the column.

Integrated Guard Column Design Guard Holder Cartridge with Integrated Guard Column A

Guard columns are designed with relatively small total volumes and C HPLC nalytical minimal dead volumes so that they minimize band broadening. The ratio of guard column to analytical column volume should be such that adding a guard column decreases system efficiency by no more than 5–10%. Generally, a ratio of 1:10 to 1:25 is satisfactory. Although larger guard columns would have the capacity to trap impurities from more sample injections, excessive band broadening Did you know... would result. Waters offers two configurations for guard columns— Syringe filters can extend the life of your analytical C and olumns the Sentry guard holders and Guard-Pak™ holder. The Sentry guard column by removing particulates from your sample. holder is available in two designs: one to be used as an integrated part of the cartridge column with reusable endfittings; the other for For more information, see page 65. use with any HPLC column. Both are installed without tools. The Guard-Pak holder is a convenient and inexpensive device since you can use prepacked Guard-Pak inserts matched to the chemistry of your analytical column. onsumables

Positioning of a Guard Column in an LC System

Solvent Delivery Injector Detector System

Guard Column HPLC Column or Inline Filter

Flow 

See page 160 for part numbers and ordering information.

147 Guard-Pak Guard-Pak Holder Inexpensive Column Protection Guard-Pak inserts prolong column life by removing contaminants from the sample and mobile phase, giving you enhanced reproducibility

and performance. They are packed with the same high performance  packings used in Waters analytical columns. Waters Guard-Pak holder is a compact, stand-alone housing for our See page 160 unique disposable Guard-Pak inserts. Installed on-line with your for part numbers and HPLC system immediately before the analytical column, the ordering information. Guard-Pak holder and inserts protect analytical LC columns against

the gradual accumulation of particulates and chemical contaminants

originating from the sample. 

See page 160 for part numbers and ordering information.

Waters Sentry Guard Holders and Sentry Guard Columns Guard-Pak Inserts Guard columns are widely used as a cost effective means of prolonging onsumables HPLC column life. Guard columns protect the column from particulate Guard-Pak inserts protect your analytical columns from damage due matter and chemical contaminants. to irreversibly adsorbed chemical contaminants or particulates. The result is enhanced chromatographic reproducibility and improved The high-performance Sentry guard column was developed for customers column lifetime. The inexpensive, disposable plastic inserts are needing the increased capacity required for biological samples and/ prepacked with analytical grade 4, 5, and 10 µm packing materials or temperature control. When placed in series with an analytical and sealed with 2 µm filters. Unlike hand-packed guard columns, column, Waters guard column increases overall efficiency. This Guard-Pak inserts have a consistent, densely packed bed and are olumns and C efficiency increasing phenomenon was observed when the Sentry guard designed to eliminate voids thus ensuring column-to-column column was used with both short (100 mm) and standard length reproducibility.

analytical columns. The Sentry guard column gives you double the capacity of traditional  guard columns so that you need to change the column less frequently. For Guard-Pak column part In addition to extending the productive lifetime of your column, the numbers, see column part Sentry guard column enhances the reproducibility giving you consistent, number tables starting on reliable, reproducible results time after time. nalytical HPLC C page 160 A Sentry guard columns contain the same high-performance packing materials that are used in all Waters analytical columns. These convenient, easy-to-use, disposable guard columns are an economical means of protecting your more expensive analytical column. You can change the guard column in seconds. Two holder designs are offered, one to be used as an integrated part of the Waters cartridge column with reusable endfittings, the other for use with any HPLC column. Both allow Sentry guard columns to be replaced without tools.

148 www.waters.com/order Shodex RSpak Polymer Reversed-Phase Columns

Shodex RSpak® columns are packed with porous polymeric particles that Comparison of Basic Compounds on Five Different are stable at a pH range of 2–12 and have a different selectivity from RSpak Columns and C18 Based Column silica-based columns. The wide pH range makes the chromatography Sample: Local anesthetics of basic compounds much easier because it is possible to chromatograph Solvent: 44% acetonitrile, 56% water, 1% ammonium hydroxide (pH 11) at high pH without ion-pairing salts. Waters offer six different types Flow Rate: 3 mL/min Detection: UV @ 220 nm of polymeric packings. One of these, the DS-613, is based on a styrene divinylbenzene copolymer which is similar to ‘conventional’ 12 polymer-based columns and works well with more hydrophobic samples requiring high concentrations of organic modifiers. 3 The DE-613 polymethacrylate and the family of functionalized packings A: DE-613 (D18-613 and D4-613) are naturally more hydrophilic than the DS-613 packing and function well in mobile phases containing high 1 concentrations of water. The least hydrophobic of these packings is 2 the DE-613 followed by the D4-613 and the longer carbon chain length derivative columns. The DC-613 is a cation exchanger and has a unique selectivity (mixed chromatographic mode ion-exchange and 3 B: DS-613

reversed-phase partition) for weakly cationic species. A HPLC C HPLC nalytical 1 RSpak D Series 2 3 E: C18

Column Type Base Polymer Functional Group DS-613 Polystyrene None 18 min DS-613 Polymethacrylate None Above is a comparison of the separation of three local anesthetics using RSpak columns DS-613 Polystyrene Sulfonated DE-613, DS-613, D18-613 and D4-613. Mobile phase contains ammonium hydroxide at

pH 11. The compounds separated are lidocaine, procaine and tetracaine. The pK values of

the anesthetics are greater than 9.0, a mobile phase with a high pH is therefore needed to C and olumns  deprotonate these compounds. Once deprotonated, they are easily separated by a reversed- phase mechanism. Eluents are simple and easier to reproduce. See page 163 for part numbers and ordering information. onsumables

Choosing the Appropriate RSpak Column Proteins and Peptides for your Separation Column: RSpak D18-613, If you had no prior experience with polymeric reversed-phase (PRP) 6 x 150 mm Part Number: WAT036576 packing materials, the efficient 6 µm polymethaceylate based RSpak Mobile Phase A: 0.05% TFA: DE-613 column would be a good starting point for most applications. acetonitrile (80:20) Mobile Phase B: 0.05% TFA: This column exhibits a mid-range hydrophobicity index equivalent to a acetonitrile (30:70) Linear gradient A to B C8 bonded silica phase. in 30 min Flow Rate: 1 mL/min Should increased retention be required for the analyte of interest, Detection: UV @ 220 nm the more hydrophobic SDVB based DS-613 column can be used. Alternatively, decreased retention is observed with the functionalized Compounds: 1. Bradykinin methacrylate based packings such as D18-613 and D4-613 and 2. Neurotensin should be used if analytes are very hydrophobic. 3. Substance P 4. Insulin 5. Insulin B chain 6. Lysozyme 7. Myoglobin

30 min

149 Paired-Ion Chromatography (PIC Reagents)  See page 179 An easy solution to the problem of analyzing samples containing ionic species, Waters PIC® reagents allow ionic compounds to be separated by reversed-phase chromatography while eliminating problems for part numbers and ordering information. of precise pH and temperature control, reproducibility, and short column life associated with ion exchange. PIC reagents versatility can save time by allowing simultaneous assay of acids, bases, amphoteric, and neutral compounds.

Waters PIC reagents are: Compound or Class PIC Reagent Compound or Class PIC Reagent Purified and quality controlled by liquid chromatography to Alkaloids B Vitamins B5/B7 guarantee consistent results Cephaeline B Calcium Leucovorin A Preformulated for ease of use Emetine B Buffered to an appropriate pH for most acidic and basic compounds Ergot alkaloids B FD&C Dyes A The accompanying table lists some typical compounds which can be Analgesics/Antipyretics Folic Acid A chromatographed using paired-ion chromatography. A µBondapak C18 column with a water/methanol solvent system was used in all cases. APC A Germicides Aspirin A Use Low UV PIC to analyze ionic compounds at low UV (200 nm). Benzalkonium Codeine B B Low UV PIC is specially formulated to eliminate background interference Chloride Indomethacin A in the low UV range and provides: Cetylpyridinium B Meperidine B Methods development opportunities for ion pairs which do not have a chromophore in the working range of regular PIC reagents Methadone B Imides A Morphine B Increased sensitivity for compounds ordinarily used with existing Methotrexate A PIC reagents Propoxyphene B

onsumables Potential for selectivity changes due to alternative buffers as Petroleum Sulfonates A Antibiotics compared to regular PIC reagents Cephalosporins A or B Surfactants Use PIC A for rapid separation of acids. FD&C sulfonic acid dyes are Penicillins A or B Quaternary amines B classic examples of compounds that are conventionally separated by Sulfonates A Antihistamines ion-exchange chromatography. Using PIC Reagent A and reversed- phase chromatography, the sulfonic acid dyes and their impurities Chlorpheniramine B Sympathomimetics olumns and C can be separated easier, faster, and more reliably. Diphenhydramine B (Decongestants) Amphetamine B Use PIC B for rapid separation of bases. Quaternary amines, another Methapyrilene B Ephedrine B class of compounds that is usually separated by ion-exchange chroma- Phenimdamine B tography, can be more easily separated by paired-ion chromatography Pheniramine B Epinephrine B employing one of the PIC Reagent B series. Pyrilamine B Isoproterenol B Use Reagent PIC D-4 as a mobile-phase modifier when chromatographing Tripelennamine B Naphazoline B basic pharmaceutical compounds to improve peak shape and reduce Nylidrin B nalytical HPLC C

A retention times. Antitussives B Phenylephrine B Phenylpropanolamine B FD&C Dyes Cough Syrup Choice of B Reagent will depend upon the mixture to be separated. B7 is the recommended Column: µBondapak C18, 3.9 x 300 mm, 10 µm initial choice. Part Number: WAT027324 Solvent: a) water with PIC Reagent A b) methanol with PIC Reagent A 40% B–55% B Sample: 80 µL cough syrup (diluted 1/1 with water) Flow Rate: 2 mL/min Detection: UV @ 254 nm Gradient Profile

Red 40

Yellow 6

PIC techniques can be used to determine trace impurities in bulk dyes or to quantitate dyes in a complex mixture, such as a cough syrup.

150 www.waters.com/order Sugar and Carbohydrate Analysis  See page 178 Carefully controlling the content of commercial foods and beverages is the fundamental task of any manufacturer. Quality control is of for part numbers and ordering information. premier importance, not only for compliance with regulatory agencies, but for customer satisfaction as well. The primary ingredients of foods and beverages, which must be Selecting the Column analyzed and quantitated, are sugars and carbohydrates, fatty acids, The table on page 153 lists the various columns along with the triglycerides and cholesterol, pesticides, organic acids, alcohols, associated separation mode, typical mobile phases, and expected caffeine, and other additives. order of carbohydrate elution. Waters offers a range of columns for analysis of each of these classes For Shodex columns, the name of a particular column contains all the of compounds. pertinent information necessary to define the column. These factors Sugars constitute a very important class of compounds which are are listed in the second table. ubiquitous in nature. They are found in food, microorganism, and paper pulp, and are involved in many biochemical interactions. Sugars present Selecting the Separation Mode a challenge for separations because of the small differences in their There are many different modes of chromatography used to characterize chemical and physical properties. In order to exploit these minor carbohydrates and many different columns from which to choose. differences and effect a separation, several modes of chromatography Before choosing a column, an understanding of the types of columns exist for the analysis of sugars. that are available and the mode of chromatography that can be used A HPLC C HPLC nalytical to exploit the differences between sugars would be beneficial.

Ion Exchange This mode provides selectivity to separate mono- and some disaccharides, Did you know... as well as sugar alcohols and low molecular weight alcohols, by hydroxyl Waters offers a wide range of applications to help you coordination to the metal cation. Ion exchange uses sulfonated styrene with your analyses. divinylbenzene resins in lead form (Shodex SP-0810) or calcium

Search by compound name, family, or class of compound at form (Shodex SC-1011, or our Sugar-Pak I). The mobile phase is C and olumns water at 80–90 ˚C. www.waters.com/applications Partition Partition chromatography is best suited for low molecular weight sugars such as mono-, di-, and trisaccharides. This mode uses a covalently bonded amino packing in a column such as our Carbohydrate Analysis onsumables column, or a dynamically-coated silica cartridge employing silica amine modification (SAM™) reagents such as SAM I. Typical mobile phases range from 65–85% acetonitrile in water, and the separations are carried out at temperatures ranging from ambient to 70 ˚C.

Reversed Phase Reversed phase is best suited to the separation of mixtures containing monosaccharides and oligosaccharides up to a degree of polymerization (DP) of 8 as applies to monitoring acid or enzyme hydrolysates of large oligosaccharides. This mode is also amenable for the separation of derivatized sugars.

Size Exclusion Gel permeation chromatography (GPC) is used to determine the molecular weight distribution of food polymers such as starches and gums. Waters Ultrahydrogel columns are well suited for this application. Shodex KS Series (7 µm) and robust S Series (10 µm) packings are based on a sulfonated styrene divinylbenzene polymer. Designed to analyze and characterize water-soluble polymers, they are particularly well suited for mono-, di- and polysaccharides. The KS-800 Series is also useful for separation of intermediate and small DP sugars.

151 Typical Applications for Sugar and Carbohydrate Columns

SH-1011, IC-Pak Cartridge/ Carbohydrate SAM I Reagent with Sugar-Pak I, SC-1011, Ion-Exclusion Dextro-Pak™ KS-800 series Column Analysis Column Silica Cartridge SP-0810 Fast Fruit Juice Ion exchange/ Ion exchange/ Mode Partition Partition Reversed phase Size exclusion size exclusion size exclusion 65–85% acetonitrile/ 70–80% acetonitrile/ 0.01 N phosphoric Eluent water ambient to water 0.1% SAM I Water 75–95 ˚C Water ambient acid 50–60 ˚C — 70 ˚C ambient Mono-, di- and trisac- Mono-, di-, Mono- through Mono-, di- and Sugar acids, sugar Hydrolysed syrups, Application charides up to DP 8 oligosaccharides oligosaccharides trisaccharides alcohols, organic acids derivatized sugars Sugars and sugar alcohols and sugar alcohols such as syrups

Elution Largest and most

Smallest elute first Smallest elute first Largest elute first Smallest elute first Largest elute first Order acidic elute first

 

See page 178 See page 294 for part numbers and for part numbers and ordering information. ordering information.

Guide to Shodex Sugar Columns onsumables

S C 18 2 1 Type of column Cation % Cross Linkage Pore Size 0 - Gel Type S = sugar H = H+ 1 = 20Å 1 - Semimacropore gel C = Ca2+ 2 = 50Å 2 - Permanent pore gel P = Pb2+ 3 = 100Å olumns and C Z = Zn2+ 4 = 500Å 5 = 1000Å Example: S C 10 1 1 Sugar column Ca2+ 10% cross linkage 20Å pore size semimacropore gel nalytical HPLC C A

152 www.waters.com/order Retention Times for Sugar and Carbohydrates

Column Type Carbohydrate Analysis Sugar-Pak Dextro-Pak Silica-Pak™ KS-801 SC-1011 SP-0810 SH-1011 Column Temp: Ambient 90 ˚C Ambient Ambient SAM I Reagent 75 ˚C 80 ˚C 80 ˚C 60 ˚C Mobile Phase: 80/20 Acetonitrile/Water Water Water 75/25 Acetonitrile/ Water Water Water Water Water Flow Rate: 2 mL/min 0.5 mL/min 1 mL/min 4 mL/min 1 mL/min 1 mL/min 1 mL/min 1 mL/min

Compound Approximate Elution Time in Minutes 2-Deoxygalactose — — — — — 8.4 10.7 7.9 2-Deoxyribose — — — — — 9.2 10.8 8.6 2-Deoxy-glucose 3.9 8.8 3.4 1.5 — 7.7 9.3 7.4 Adonitol (Ribitol) 4.8 11.0 2.9 1.8 — 9.8 13.5 7.7 Arabinose 4.8 10.4 3.0 1.8 8.3 9.0 11.1 7.5 Arabitol — — — — — 11.1 16.5 8.0 Butanol-1 N/R 18.8 N/R — — — — — Cellobiose 15.3 6.7 3.7 4.1 5.7 5.4 7.9 5.5 Deuterium Oxide — — — — 11.1 — — — Digitose — — — — — 9.3 10.6 — Digitoxose — — — — — — — 8.4 Dulcitol — — — — — 12.7 20.8 7.6 Ethylene Glycol N/R 12.6 3.5 1.0 9.3 — — — Ethanol N/R 12.8 6.8 — 10.0 — — —

Fructose 5.7 10.1 2.9 2.0 7.7 9.0 11.5 7.1 A Fucose 4.2 10.4 4.1 1.6 8.1 8.9 11.2 7.8 C HPLC nalytical Galactose 7.7 9.4 2.8 2.5 7.6 8.2 10.4 6.9 Glucoheptose — — — — — 9.6 — 6.6 Glucose 7.3 8.5 2.8 2.4 7.2 7.6 9.1 6.6 Glucose-6-phosphate N/R 4.8 2.1 — — — — — Glycerol N/R 12.1 3.2 1.2 8.5 — — — Inositol — — — — — 9.0 13.6 7.0 Isomaltitol — — — — — 8.7 15.3 — Isomaltose — — — — 5.9 6.4 8.1 5.6 C and olumns Lactose 18.0 7.1 3.0/3.2* 4.8 5.9 6.6 8.6 5.8 Lactulose 16.3 7.6 3.8 4.0 — 7.1 9.9 — Lyxose 4.1 10.7 3.0 — — 9.3 11.4 7.4 Maltitol — — — — 6.0 — 13.2 6.1 Maltoheptaose — — — — — — 6.6 4.6 Maltohexaose — — — — — 5.4 6.8 4.6 Maltopentaose — — — — 4.7 5.3 7.2 4.7 Maltose 15.7 6.8 3.5 4.1 5.9 5.5 8.3 5.6 onsumables Maltotetraose — — — — — 5.8 7.5 4.9 Maltotriose N/R 6.0 4.4/4.7* 7.8 5.3 6.1 7.9 5.1 Maltotriol — — — — — 5.9 11.3 5.4 Mannitol 7.0 12.2 2.9 2.2 7.2 11.1 16.5 7.4 Mannose 6.4 9.6 2.9 2.2 7.6 8.3 11.4 6.9 Melibiose 21.6 7.0 3.1 5.3 5.9 6.4 8.7 5.6 Melizitose — — — — 5.1 6.0 7.3 5.1 meso-Erythritol — — — — — 10.1 13.3 8.4 Methanol N/R 12.5 3.9 — — — — — myo-Inositol — — — — — — 13.5 7.0 Pannose — 5.9 5.0 — — — — — Pentaerythritol — — — — — 9.8 11.5 9.1 Propanol-2 N/R 12.6 N/R — — — — — Raffinose N/R 6.0 7.0 7.5 5.2 6.1 7.5 5.1 Rhamnose — 9.5 3.3 — 7.3 8.3 10.3 7.3 Ribose 3.6 14.9 3.1 1.4 9.1 13.5 19.9 7.9 Sorbitol 6.9 14.2 2.9 2.2 7.5 13.2 22.3 7.6 Sorbose 5.6 9.4 2.8 — 7.4 8.2 10.2 6.7 Stachyose N/R 5.4 6.3 17.5 4.8 5.7 7.1 4.9 Sucrose 11.9 6.8 4.4 3.4 5.8 6.4 8.0 — Trehalose — — — — 5.7 6.4 8.1 5.6 Turanose — — — — — 6.6 8.5 5.8 Xylitol 5.1 14.3 2.9 1.8 8.0 13.0 20.5 8.2 Xylose 4.3 9.3 2.9 1.6 7.7 8.1 9.7 7.0 Xylulose 3.3 10.6 3.1 — — — — —

N/R - Not recommended under these conditions due to insufficient or excessive retention. * - Elution times of anomers Blank - No data exists

153 Application-Specific Columns  See page 178 High-Performance Carbohydrate Analysis Cartridge Column for part numbers and ordering information. Waters High-Performance Carbohydrate cartridge column with reusable endfittings is based on a 4 µm, spherical silica. This column was developed to separate 5 monosaccharide and disaccharides with base line resolutions in less than 12 minutes. The High-Performance Carbohydrate cartridge column can be run at 1.4 mL/min to achieve 12-minute separations and is available in a 4.6 mm ID x 250 mm cartridge column. This column dimension was found to be the optimum for speed, resolution and longevity. The prepacked, disposable cartridge column requires reusable endfittings which can be purchased separately. When you change columns, just replace the cartridge and use the endfittings again. You can increase the lifetime of the column by using a Sentry guard column. Contaminants are removed from the sample and mobile phase before they reach the column.

Fatty Acid Analysis Pesticide Analysis Fatty acid analysis can be carried out by the reversed-phase mode. When used with the Waters Carbamate Analysis system, the Carbamate The Free Fatty Acid HP column uses a phenyl-bonded packing and a Analysis column provides a guaranteed analysis of the carbamate simple isocratic elution method to separate free fatty acids based pesticides that exceeds the sensitivity required by AOAC Method 985.23. on carbon chain length and degree of saturation. The short column The baseline resolution and high sensitivity of this separation, coupled dimension (3.9 x 150 mm) significantly reduces analysis time and with the optimized system configuration, provide state-of-the-art increases sensitivity. Typical applications include free fatty acids, analysis of carbamates. The separation of eleven carbamate pesticides fatty acid derivatives, and some fatty amides, such as saponified and carbamate metabolites is accomplished in 20 minutes.

onsumables coconut oil, tall oils, fatty alcohols, and fatty acid isomers. Straight chain saturated acids elute in order of increasing carbon number. Carbamate Analysis Unsaturated acids elute before the analgous saturated compound. Column: Carbamate Analysis Compounds: For a given carbon number and chain configuration, the greater 3.9 x 150 mm, 4 µm 1. Aldicarb sulfoxide Part Number: WAT035577 2. Aldicarb sulfone the unsaturation, the earlier the elution. Mobile Phase: Water/methanol/acetonitrile 3. Oxamyl olumns and C complex gradient 4. Methomyl Flow Rate: 1.5 mL/min 5. 3-Hydroxycarbofuran Sample: 10 ng of each analyte on column 6. Aldicarb Fatty Acid Standards Injection Volume: 400 µL 7. Propoxur Post Column Addition: OPA/sodium hydroxide @ 0.5 mL/min 8. Carbofuran Column: Free Fatty Acid HP, Detection: Model 470 Fluorescence detector 9. Carbaryl 3.9 x 150 mm, 4 µm Excitation: 339 nm lambda 10. 1-Naphthol Part Number: WAT011690 Emission: 445 nm lambda 11. Methiocarb Mobile Phase: Acetonitrile: THF: water 45:20:35 5 Flow Rate: 1.5 mL/min Detection: RI 9 nalytical HPLC C

A Compounds: 1. Capric acid 1 2. Lauric acid 2 7 3. Myristic acid 6 8 4. Palmitic acid 3 11 5. Stearic acid 4 6. Nonadecanoic acid 7. Arachidic acid 10 8. Heneicosanoic acid 1 9. Behenic acid 2 3 4 5 6 7 89

5 min 20 min

 

See page 178 See page 178 for part numbers and for part numbers and ordering information. ordering information.

154 www.waters.com/order Waters PAH Columns  See page 178 Improve Analysis of PAH Compounds for part numbers and ordering information. Polynuclear aromatic hydrocarbons (PAHs) are among the most frequently monitored environmental contaminants. Standard and official methods for the analysis of PAHs are found in compendia for air, drinking water, wastewater, solid waste, and food analysis.1 Many of these methods specify HPLC, usually with UV and fluorescence detection, as the recommended analytical procedure. Waters PAH columns are optimized for the HPLC analysis of PAHs. The chromatogram (bottom left) shows 16 PAH compounds, listed as target pollutants by the U.S. EPA. The Waters PAH columns achieve baseline resolution and excellent peak symmetry for all 16 target analytes in less than 25 minutes, while employing a simple water: acetonitrile binary gradient. The resolving power of the PAH columns provides superior peak identification and quantitation for PAHs. Florida Administrative Code 17.700 includes 2 additional compounds (1-methyl naphthalene and 2-methyl naphthalene) in addition to the

16 compound EPA 610 mix that we currently use to show the proficiency of Waters instrumentation to analyze PAH compounds (bottom right). A The new Waters PAH columns resolve these two compounds along with the other 16. C HPLC nalytical Waters PAH columns come in seven different dimensions (including a capillary format) and two particle sizes. Each column comes with a complete certificate of analysis backed by a world-class ISO 9002 registered documentation trail.

Reference: 1 AOAC 973.30; Deutsche DIN TVO; UK ISBN 0 11 & 752032 2; U.S. EPA Methods TO-13, 550 & 550.1, 610, 8310

PAH Analysis using Waters PAH Columns PAH Analysis According to Florida Administrative Code 17.700 C and olumns

Column: Waters PAH C18, 4.6 x 250 mm, Compounds: Column: Waters PAH C18, 4.6 x 250 mm, Compounds: 5 µm at 27 °C 1. Naphthalene 20 ppm 5 µm at 27 °C 1. Naphthalene 20 ppm Part Number: 186001265 2. Acenaphthylene 40 ppm Part Number: 186001265 2. Acenaphthylene 40 ppm System: Waters Alliance with 2996 3. Acenaphthene 20 ppm Eluent A: Water 3. 1-methyl naphthalene* 25 ppm Photodiode Array detector 4. Fluorene 4 ppm Eluent B: Acetonitrile 4. 2-methyl naphthalene* 25 ppm Eluent A: Water 5. Phenanthrene 2 ppm Gradient: 60% B to 100% B using curve 9 5. Acenaphthene 20 ppm Eluent B: Acetonitrile 6. Anthracene 2 ppm in 12 minutes, hold 11 minutes, 6. Fluorene 4 ppm onsumables Gradient: 60% B to 100% B using curve 9 7. Fluoranthene 4 ppm back to initial conditions 7. Phenanthrene 2 ppm in 12 minutes, hold 11 minutes, 8. Pyrene 2 ppm Flow Rate: 1.2 mL/min 8. Anthracene 2 ppm back to initial conditions 9. Benzo(a)anthracene 2 ppm Injection: 20 µL 9. Fluoranthene 4 ppm Flow Rate: 1.2 mL/min 10. Chrysene 2 ppm Sample: EPA-610 mixture plus 10. Pyrene 2 ppm Injection: 20 µL 11. Benzo(b)fluoranthene 4 ppm two compounds 11. Benzo(a)anthracene 2 ppm Sample: EPA-610 mixture 12. Benzo(k)fluoranthene 2 ppm 12. Chrysene 4 ppm 13. Benzo(a)pyrene 2 ppm 13. Benzo(b)fluoranthene 4 ppm 14. Dibenzo(a,h)anthracene 4 ppm 14. Benzo(k)fluoranthene 2 ppm 15. Benzo(g,h,I)perylene 4 ppm 15. Benzo(a)pyrene 2 ppm UV @ 254 nm 16. Indeno(1,2,3-cd)pyrene 2 ppm UV @ 254 nm 16. Dibenzo(a,h)anthracene 4 ppm 17. Benzo(g,h,I)perylene 4 ppm 6 18. Indeno(1,2,3-cd)pyrene 2 ppm

10 12

2 9 1

5 7 8 13 4 7 13 15 12 2 1 11 4* 3 8 15 16 3* 6 9 10 14 14 17 5 16 18

6810 12 14 16 18 20 22 24 min 579 11 13 15 17 19 21 23 25 min

155 Triglycerides and Cholesterol Analysis

Non-aqueous, reversed-phase chromatography is the best method of Cholesterol from Butter separating triglycerides into classes based on carbon number and degree of unsaturation. This method aids in measuring oil degradation Column: Ultrastyragel 100Å and 500Å, 7.8 x 300 mm, 5 µm Part Numbers: WAT085500 (100Å) & WAT010571 (500Å) and adulteration. Waters Nova-Pak C18 column is an excellent choice Mobile Phase: Chloroform for this separation due to its high efficiency and carbon surface area. Flow Rate: 1.0 mL/min Sample: A: cholesterol standard, 0.125% It can also be used for cholesterol determination from samples, such B: butter (1/10 w/v) = 0.24% in as meat following saponification. undiluted sample Injection Volume: 50 µL The analysis of high levels of cholesterol in samples like butter Detection: RI 410 Refractometer @ 4x and antioxidants in oil, without saponification, can be effectively

performed by size exclusion using Waters Ultrastyragel™ columns. A 

See page 294 B for part numbers and ordering information.

22 min

Fermentation Analysis, Organic Acids, Alcohols, and Carbohydrates

onsumables The ion-exclusion mode is ideally suited for the separation of KC-811 Column Retention Chart for Organic Acids monosaccharides, organic acids, or sugar acids. The column packings Sample Retention Time Sample Retention Time are sulfonated styrene divinylbenzene resins in the hydrogen form ß- Hydroxy- (IC-Pak Ion Exclusion or SH-1011), and the mobile phase is a dilute Oxalic Acid 5.20 8.60 propionic Acid acid such as 0.01 N phosphoric acid at 50–60 ˚C. Maleic Acid 5.80 D-Glucuronic Acid 8.65 Using this mode, the Fast Fruit Juice column can effectively separate a-Ketoglutaric Acid 5.90 Fumaric Acid 8.95

olumns and C glycerol, acetic acid, and ethanol in grape or other fruit juices. The Citric Acid 6.20 Formic Acid 9.20 degree of microbial defect, the extent of natural fermentation in grapes and the amount of sulfite present in various foods and beverages Tartaric Acid 6.55 Acetic Acid 9.80 can also be analyzed with the Fast Fruit Juice column. The IC-Pak Pyruvic Acid 6.65 Adipic Acid 9.80 ion-exclusion column can separate a wide range of organic acids while trans-Aconitic Acid 6.95 Levulinic Acid 10.00 the Shodex SH column separates acids as well as larger carbohydrates. Glyoxylic Acid 7.00 Mesaconic Acid 10.40 The analysis of alcohols and organic acids is important as these Malic Acid 7.05 Pyroglutamic Acid 10.70 compounds typically play a role in determining the flavor characteristics Malonic Acid 7.07 Propionic Acid 11.25 nalytical HPLC C

A of beverages such as wine, beer and some spirits. The presence of Citraconic Acid 7.20 Acrylic Acid 11.60 alcohols in fruit juices can indicate product deterioration. The Shodex Succinic Acid 8.00 Pivalic Acid 14.05 KC-811 column is packed with a sulfonated rigid styrene divinylbenzene copolymer providing high efficiency separations of low molecular weight Glycolic Acid 8.40 Methacrylic Acid 14.10 organic acids and water soluble organics such as alcohols, aldehydes, Itaconic 8.50 trans-Crotonic Acid 15.65 and nitriles. The column provides ion exclusion and reversed-phase Lactic Acid 8.60

mode of chromatography. Typical mobile phases are aqueous Eluent: Water with 0.1% phosphoric acid, Temperature: 60 ˚C, Flow Rate: 1 mL/min

solutions containing 1% phosphoric, acetic or perchloric acid run

1 mL/min at 45–80 ˚C. 

See page 178 for part numbers and ordering information.

156 www.waters.com/order Ion Analysis  Waters offers an array of products for ion chromatography as well as innovative capillary electrophoresis See page 178 products for ion analysis. In ion chromatography, anions and cations are typically measured in two separate for part numbers and analyses. The columns offered for each type of analysis are briefly described below. ordering information.

Waters IC-Pak resin-based columns allow you to analyze a full range IC-Pak Anion Column of ions from numerous sample matrices, both simple and complex. They offer an exceptional linear loading range of less than 1.0 ppb Column: IC-Pak Anion, 4.6 x 50 mm, 10 µm Compounds: to greater than 400 ppm without dilution and without pH limitations Part Number: WAT007355 1. Fluoride 1 ppm Sample: Seven anion standard 2. Bicarbonate — on eluent or sample. The flexibility exists for accurate and reproducible Eluent: Borate/gluconate 3. Chloride 2 ppm anion and cation analyses at all concentration levels. The IC-Pak series Detection: Waters 431 Conductivity detector 4. Nitrite 4 ppm 5. Bromide 4 ppm of resin-based columns shares the same chemistry and gives you 6. Nitrate 4 ppm 7. Phosphate 6 ppm identical elution order profiles. 3 8. Sulfate 5 ppm

Anion Analysis 1 6 The IC-Pak Anion series of columns is used for the analysis of inorganic 5 anions, while the IC-Pak Ion-Exclusion columns are used for organic 7 8 acid analysis. 2

The silica-based IC-Pak C M/D column allows a simultaneous analysis A of monovalent and divalent cations under isocratic conditions. This C HPLC nalytical technology provides the analytical chemist with the most sensitive 15 min means of analyzing monovalent cations such as Na+, K+, Li+, and NH4+. The IC-Pak Anion column is a configuration of 10 µm anion-exchange packing material and a short column length which makes this the column of choice for rapid routine analyses. The IC-Pak Cation column is packed with 10 µm sulfonated styrene divinylbenzene particles. Monovalent and divalent cations may be analyzed in two separate runs. Different mobile-phase compositions are used for the two analyses. IC-Pak C M/D - Cation Column C and olumns Method: Waters IC-Method C-207 Column: Waters IC-Pak C M/D, 1 Cation Analysis 4 3.9 x 150 mm 2 3 The selection of a cation analysis column depends on the type of Part Number: WAT036570 5 Eluent: 0.1 mM EDTA/3.0 mM cation being measured. The IC-Pak C M/D column separates alkali and nitric acid 6 Flow Rate: 1 mL/min alkaline earths more efficiently than the traditional IC-Pak Cation Injection: 100 µL column. The IC-Pak C M/D column also separates ethanolamine- Detection: Conductivity onsumables related organic cations. Transition metals can be separated using a Compounds: Delta-Pak C18 column. The lanthanide series can be separated using 1. Lithium 0.54 ppm 2. Sodium 1.0 ppm Resolve C18 (a nonendcapped C18 material). Finally, metallocyanides 3. Ammonium 2.0 ppm can be analyzed on a Nova-Pak C18 column. 4. Potassium 4.0 ppm 5. Magnesium 2.0 ppm

6. Calcium 3.0 ppm 14 min

Did you know... IC-Pak Anion columns are available in HC (high-capacity) and HR (high-resolution) formats. For ordering information, see page 178.

157 Transition Metal Analysis Lanthanide Analysis

Transition metals can be separated on a dynamically coated C18 Lanthanide Analysis using a Resolve C18 Radial-Pak Cartridge column and detected at low ppb levels using post-column addition

of PAR reagent with UV detection. This method provides excellent Column: Resolve C18 Radial-Pak Cartridge, 8 x 100 mm, 5 µm resolution with good selectivity and analysis time. Part Number: WAT084624 Sample: Lanthanides Injection: 10 µL Eluent A: 10 mM sodium octanesulfonate,100 mM hydroxyisobutyric acid, pH 3.8 Transition Metal Analysis using Post-Column Derivatization Eluent B: 10 mM sodium octanesulfonate, 400 mM hydroxyisobutyric acid, pH 3.8 Flow Rate: 2.0 mL/min Post-Column Column: Delta-Pak C18, 100Å, 3.9 x 150 mm, 5 µm Compounds: 9 Reagent: 0.13 mM arsenazo III, 10 mM urea, 7 8 10 Part Number: WAT011795 1. Cu 0.5 ppm 62 mM acetic acid at 1 mL/min Injection: 100 µL 2. Pb 1.0 ppm Detection: UV @ 658 nm 6 Flow Rate: 0.8 mL/min 3. Zn 0.5 ppm 11 12 13 Eluent: 2 mM sodium octanesulfonate, 4. Ni 1.0 ppm 35 mM sodium tartrate, 5% acetonitrile, 5. Co 0.5 ppm adjust to pH 3.65 with sodium hydroxide 6. Cd 1.0 ppm Post-Column Reagent: 0.2 mM PAR [4-(2-pyridylazo)resorcinol], 7. Fe(II) 2.0 ppm Compounds: 2 ppm of each 1 M acetic acid, 3 M ammonium hydroxide, 8. Mn 1.0 ppm 5 0.5 mL/min 1. Lu 8. Gd 8 2. Yb 9. Eu 3. Tm 10. Sm 14 1 3 4. E 11. Nd 5 5. Ho 12. Pr 6. Dy 13. Ce 4 7. Tb 14. La

4 7 3 2 6 2

1 onsumables

20 min

17 min

 

See page 266 See page 200 for part numbers and for part numbers and

olumns and C ordering information. ordering information.

Ion-Exclusion Columns

nalytical HPLC C Waters IC-Pak Ion Exclusion columns are used for the analysis of weak acid anions such A

as fluoride and short chain organic acids from formate to butyrate. 

See page 178 for part numbers and ordering information.

Did you know... Waters Analytical Standards and Reagents offers a comprehensive portfolio of application-specific standards to support your system. For more information, see page 300.

158 www.waters.com/order Gas-Chromatography Packings  Waters versatile PoraPak gas-chromatography column packing materials simplify the analysis of a wide See page 176 variety of complex compounds from atmospheric gases to organics. Consisting of polymer beads, these for part numbers and unique packings are chemically and physically stable and have consistent particle size, porosity, and ordering information. surface area to guarantee analytical reproducibility. They also provide unequaled separation capability with high resolution and low, constant retention volumes. Versatility for Specialty Applications To optimize separation of even the most complex matrices, several physical and chemical variations are available within the PoraPak packing materials. Special characteristics of Waters unique GC packings include: Fast analysis with compounds eluting in distinctive bands with no tailing The ability to sustain elevated temperatures permitting temperature programming without adverse effects to retention, reproducibility, and column life The ability to accommodate large sample loads required for preparative and trace analysis while maintaining characteristically high column efficiency

GC PoraPak Porous Polymer Packing

Type Polarity Surface Area (m2/g) Density (g/cm3) Single Temp Prog. Particle Size Mesh A HPLC C HPLC nalytical 50–80 P Nonpolar 100–200 0.26 250 ˚C 80–100 100–120 50–80 PS Nonpolar 100–200 0.26 250 ˚C 80–100 100–120

50–80 C and olumns Slightly nonpolar Q 500–600 0.34 250 ˚C 80–100 to moderate 100–120 50–80 Slightly nonpolar QS 500–600 0.34 250 ˚C 80–100 to moderate 100–120 onsumables 50–80 Moderate R polar monomer: 450–600 0.32 250 ˚C 80–100 vinyl pyrrolidone 100–120 50–80 Moderate S polar monomer: 300–450 0.35 250 ˚C 80–100 vinyl pyridine 100–120 50–80 Polar N polar monomer: 250–350 0.41 190 ˚C 80–100 vinyl pyrrolidone 100–120 Highly polar 50–80 polar monomer: T 225–350 0.39 190 ˚C 80–100 ethyleneglycol dimethacrylate 100–120

Did you know... Waters has a full line of LC/GC Certified autosampler vials. For more information, see page 68.

159 >>> Product ORdering information

μBondapak/Bondapak Analytical Columns*

Dimension Type Particle Size Pore Size Qty. µBondapak C18 µBondapak CN µBondapak NH2 µBondapak Phenyl 2.1 x 300 mm Column 10 µm 125Å 1/pk WAT086609 — — — 3.9 x 20 mm Guard 10 µm 125Å 2/pk WAT0444804 WAT0468554 WAT0468654 WAT0468504 3.9 x 150 mm Column 10 µm 125Å 1/pk WAT086684 WAT086688 WAT086680 3.9 x 300 mm Column 10 µm 125Å 1/pk WAT027324 WAT084040 WAT027198 4.6 x 150 mm Column 10 µm 125Å 1/pk WAT044370 — — — 4.6 x 300 mm Column 10 µm 125Å 1/pk 186000925 — — — — Guard-Pak Inserts 10 µm 125Å 10/pk WAT08807013 WAT02675013 WAT02676013 WAT02674513 4.6 x 250 mm Cartridge 10 µm 125Å 1/pk WAT0528606 — — — 3.9 x 300 mm Column 10 µm 125Å 1/pk WAT025875 WAT084042 — —

Part No. Guard-Pak Holder WAT088141

End-Fittings* WAT037525  See page 146 for product information. 4 Requires 3.0 x 20 mm/4.6 x 20 mm Universal Sentry Guard Holder, Part No. WAT046910 6 Requires Cartridge End-Fittings, Part No. WAT037525 13 Requires Guard-Pak Holder, Part No. WAT088141 11 onsumables Requires Cartridge End-Fittings, see page 389 * Includes one pair of Reusable End-Fittings, Two C-Clips and Coupling

µPorasil/Porasil and Preparative Columns Dimension Type Particle Size Pore Size µPorasil Porasil 3.9 x 20 mm Guard, 2/pk 10 µm 125Å WAT0468603 —

olumns and C 3.9 x 150 mm Column 10 µm 125Å WAT086692 —

3.9 x 300 mm Column 10 µm 125Å WAT027477 —  See page 146 for product information. 3 Requires 2.1 x 10 mm Universal Sentry Guard Holder, Part No. WAT097958

Delta-Pak Analytical Columns

Dimension Type Particle Size Pore Size Qty. Delta-Pak C18 Delta-Pak C4 nalytical HPLC C

A 2.1 x 150 mm Column 5 µm 300Å 1/pk WAT023650 — 3.9 x 20 mm Guard 5 µm 100Å 2/pk WAT0468804 WAT0468754 3.9 x 20 mm Guard 5 µm 300Å 2/pk WAT0468904 WAT0468854 3.9 x 20 mm Guard 5 µm 100Å 10/pk WAT0368704 — 3.9 x 150 mm Column 5 µm 100Å 1/pk WAT011795* WAT011796 3.9 x 150 mm Cartridge 5 µm 300Å 10/pk WAT03687513 WAT03686513 3.9 x 150 mm Column 5 µm 300Å 1/pk WAT011793 WAT011794

Part No. Guard-Pak Holder WAT088141

Guard-Pak In-Line Filters 5/pk WAT032472  See page 146 for product information. 4 Requires 3.0 x 20 mm/4.6 x 20 mm Universal Sentry Guard Holder, Part No. WAT046910 13 Requires Guard-Pak Holder, Part No. WAT088141 *Recommended for Transition Metal Analysis, see page 158

160 www.waters.com/order Delta-Pak High Pressure Inert (HPI) Analytical Columns(PEEK) Column (non metallic) Particle Size Pore Size Dimension Part No.

Delta-Pak HPI C18 5 µm 300Å 3.9 x 150 mm WAT035571

PEEK Tubing and Fitting Kit (to minimize damage by stainless steel fittings) 1/16” OD x 0.005” ID x 5 feet (1.6 mm OD x 0.127 mm ID x 1.5 mm).

WAT022999

Kit contains 5 each of short and long compression screws, 12 poly knobs and 12 PEEK ferrules.  See page 146 for product information.

Atlantis Analytical Columns

Particle Particle Dimension Type Qty. T3 dC HILIC Silica Dimension Type Qty. T3 dC HILIC Silica Size 18 Size 18 1.0 x 50 mm Column 3 µm 1/pk 186003713 186001279 186002003 1.0 x 50 mm Column 5 µm 1/pk 186003730 186001281 186002004 1.0 x 150 mm Column 3 µm 1/pk 186003714 186001283 — 1.0 x 150 mm Column 5 µm 1/pk 186003731 186001285 — 2.1 x 10 mm Guard 3 µm 2/pk 1860037563 1860013773 1860020053 2.1 x 10 mm Guard 5 µm 2/pk 1860037593 1860013793 1860020063 Direct Direct 2.1 x 15 mm 3 µm 1/pk — 186002064 186002007 2.1 x 15 mm 5 µm 1/pk — 186002065 186002008 Connect Connect 2.1 x 20 mm Guard 3 µm 2/pk — 1860013815 — 2.1 x 20 mm Guard 5 µm 2/pk — 1860013835 — 2.1 x 20 mm IS Column 3 µm 1/pk 186003715 186002058 — 2.1 x 20 mm IS Column 5 µm 1/pk 186003732 186002059 — A

2.1 x 30 mm Column 3 µm 1/pk 186003716 186001287 186002009 2.1 x 30 mm Column 5 µm 1/pk 186003733 186001289 186002010 C HPLC nalytical 2.1 x 50 mm Column 3 µm 1/pk 186003717 186001291 186002011 2.1 x 50 mm Column 5 µm 1/pk 186003734 186001293 186002012 2.1 x 75 mm Column 3 µm 1/pk 186005652 — — 2.1 x 100 mm Column 5 µm 1/pk 186003735 186001297 186002014 2.1 x 100 mm Column 3 µm 1/pk 186003718 186001295 186002013 2.1 x 150 mm Column 5 µm 1/pk 186003736 186001301 186002016 2.1 x 150 mm Column 3 µm 1/pk 186003719 186001299 186002015 3.0 x 20 mm IS Column 5 µm 1/pk 186003737 186002061 — 3.0 x 20 mm IS Column 3 µm 1/pk 186003720 186002060 — 3.0 x 50 mm Column 5 µm 1/pk 186003738 186001391 186002018 3.0 x 50 mm Column 3 µm 1/pk 186003721 186001389 186002017 3.0 x 100 mm Column 5 µm 1/pk 186003739 186001305 186002020

3.0 x 75 mm Column 3 µm 1/pk 186005653 — — 3.0 x 150 mm Column 5 µm 1/pk 186003740 186001309 — C and olumns 3.0 x 100 mm Column 3 µm 1/pk 186003722 186001303 186002019 3.0 x 250 mm Column 5 µm 1/pk 186003741 186001311 — 3.0 x 150 mm Column 3 µm 1/pk 186003723 186001307 — 3.9 x 20 mm Guard 5 µm 2/pk 1860037604 1860013154 1860020224

3.9 x 20 mm Guard 3 µm 1860037574 1860013134 1860020214 3.9 x 50 mm Cartridge 5 µm 1/pk — 1860013874 —

3.9 x 50 mm Cartridge 3 µm 1/pk — 1860013856 — 3.9 x 100 mm Column 5 µm 1/pk — 186001395 — 3.9 x 100 mm Column 3 µm 1/pk — 186001393 — 3.9 x 150 mm Column 5 µm 1/pk — 186001319 — onsumables 3.9 x 150 mm Column 3 µm 1/pk — 186001317 — 4.6 x 20 mm Guard 5 µm 2/pk 1860037614 1860013234 1860020244 4.6 x 20 mm Guard 3 µm 2/pk 1860037584 1860013214 1860020234 4.6 x 20 mm IS Column 5 µm 1/pk 186003742 186002063 — 4.6 x 20 mm IS Column 3 µm 1/pk 186003724 186002062 — 4.6 x 30 mm Column 5 µm 1/pk 186003743 186001327 186002026 4.6 x 30 mm Column 3 µm 1/pk 186003725 186001325 186002025 4.6 x 50 mm Column 5 µm 1/pk 186003744 186001331 186002028 4.6 x 50 mm Column 3 µm 1/pk 186003726 186001329 186002027 4.6 x 75 mm Column 5 µm 1/pk 186003745 186001335 — 4.6 x 75 mm Column 3 µm 1/pk 186003727 186001333 — 4.6 x 100 mm Column 5 µm 1/pk 186003746 186001340 186002030 4.6 x 100 mm Column 3 µm 1/pk 186003728 186001337 186002029 4.6 x 150 mm Column 5 µm 1/pk 186003747 186001344 186002032

4.6 x 150 mm Column 3 µm 1/pk 186003729 186001342 186002031 4.6 x 250 mm Column 5 µm 1/pk 186003748 186001346 186002033  See page 138 for product information. 3 Requires 2.1 x 10 mm Universal Sentry Guard Holder, Part No. WAT097958 4 Requires 3.0 x 20 mm/4.6 x 20 mm Universal Sentry Guard Holder, Part No. WAT046910 5 Requires Cartridge Column Holder, Part No. 186000262 6 Requires End-Fittings, Part No. WAT037525

Atlantis Columns Method Validation Kits Each Method Validation Kit contains 3 columns, each from a different batch.

Dimension Particle Size T3 dC18 HILIC Silica 4.6 x 150 mm 3 µm 186003751 186002312 186002315 4.6 x 150 mm 5 µm 186003754 186002311 186002314

4.6 x 250 mm 5 µm 186003755 186002313 186002316  See page 138 for product information.

161 Nova-Pak Analytical Columns*

Dimension Type Particle Size Qty. Nova-Pak C8 Nova-Pak C18 Nova-Pak Phenyl Nova-Pak CN-HP Nova-Pak Silica Guard-Pak Holder Endfittings* Guard-Pak In-Line Filters Qty. Particle Size Type Dimension 2.1 x 150 mm Column 4 µm 1/pk WAT052735 WAT023655 WAT052740 — WAT052745 — — — 1/pk 4 µm Column 2.1 x 150 mm 3.9 x 20 mm Guard 4 µm 2/pk WAT0468304 WAT0443804 WAT0468354 WAT0468404 WAT0468454 — — — 2/pk 4 µm Guard 3.9 x 20 mm 3.9 x 50 mm Cartridge 4 µm 1/pk WAT0527756 WAT0527806 WAT0527906 WAT0527856 — — — — 1/pk 4 µm Cartridge 3.9 x 50 mm 3.9 x 50 mm Cartridge 4 µm 3/pk — WAT0528346 — — — — — — 3/pk 4 µm Cartridge 3.9 x 50 mm 3.9 x 75 mm Column 4 µm 1/pk WAT035877 WAT011670 WAT011675 WAT010270 WAT011680 — — — 1/pk 4 µm Column 3.9 x 75 mm 3.9 x 100 mm Cartridge 4 µm 1/pk WAT0528056 WAT0528106 WAT0528006 WAT0527956 — — — — 1/pk 4 µm Cartridge 3.9 x 100 mm 3.9 x 150 mm Cartridge 4 µm 1/pk WAT0369856 WAT0369756 WAT0369706 WAT0442436 WAT0369806 — — — 1/pk 4 µm Cartridge 3.9 x 150 mm 3.9 x 150 mm Column 4 µm 1/pk WAT035876 WAT086344 WAT010656 WAT044245 WAT010025 — — — 1/pk 4 µm Column 3.9 x 150 mm 3.9 x 150 mm Cartridge 4 µm 3/pk WAT0548706 WAT0375206 WAT0548906 WAT0444456 WAT0548806 — — — 3/pk 4 µm Cartridge 3.9 x 150 mm 3.9 x 300 mm Column 4 µm 1/pk — WAT011695 — WAT056920 — — — — 1/pk 4 µm Column 3.9 x 300 mm 4.6 x 150 mm Cartridge 4 µm 1/pk WAT0528556 WAT0528456 — WAT0444556 — — — — 1/pk 4 µm Cartridge 4.6 x 150 mm 4.6 x 150 mm Column 4 µm 1/pk — WAT044375 — — — — — 1/pk 4 µm Column 4.6 x 150 mm 4.6 x 250 mm Cartridge 4 µm 1/pk WAT0528506 WAT0528406 — WAT0444606 — — — — 1/pk 4 µm Cartridge 4.6 x 250 mm — Guard-Pak Inserts 4 µm 5/pk — — — — — — — WAT032472 5/pk 4 µm Guard-Pak Inserts —

— Guard-Pak Inserts 4 µm 10/pk WAT01522013 WAT03588013 WAT02079513 WAT02080013 WAT02079013 WAT08814113 WAT0375256 — 10/pk 4 µm Guard-Pak Inserts —

  See page 145 for product information. See page 145 for product information. 4 Requires 3.0 x 20 mm/4.6 x 20 mm Universal Sentry Guard Holder, Part No. WAT046910 6 Requires Cartridge End-Fittings, Part No. WAT037525 13 Requires Guard-Pak Holder, Part No. WAT088141 * Recommended for Triglycerides and Cholesterol Analysis, see page 156

onsumables Nova-Pak Analytical Kits Description Dimension Part No. Waters Reversed-Phase Scouting Kit 3.9 x 50 mm WAT0443606 (includes 1 each Nova-Pak C18, C8, and Phenyl Cartridge Column) Waters Method Development Scouting Kit 3.9 x 50 mm WAT0442556 (includes 1 each Nova-Pak C8, Phenyl, and CN HP Cartridge Column) Waters Method Validation Kit

3.9 x 150 mm WAT052770 olumns and C (includes 3 Nova-Pak C18 Columns representing 2 different bonded-silica bulk packing batches)  See page 145 for product information. 6 Requires Cartridge End-Fittings, Part No. WAT037525

Nova-Pak Analytical Method Validation Kit Each Method Validation Kit contains 3 columns, each from a different batch. nalytical HPLC C Description Particle Size Dimension Part No. A Method Validation Kit

4 µm 3.9 x 150 mm WAT052770

(includes 3 Nova-Pak C18 Columns)  See page 145 for product information.

Resolve Analytical Columns

Dimension Type Particle Size Pore Size Qty. C18 C8 Silica CN 3.9 x 20 mm Guard 5 µm 90Å 2/pk WAT0469153 WAT0469203 — — 3.9 x 300 mm Column 5 µm 90Å 1/pk WAT01174013 — — — 3.9 x 150 mm Column 5 µm 90Å 1/pk WAT08571113 — WAT08601613 — Guard-Pak Inserts 10 µm 90Å 10/pk WAT08582413 WAT02675513 WAT08582513 WAT08582613

Part No.

Guard-Pak Holder, Guard-Pak Inserts 10/pk WAT08814113  See page 146 for product information. 3 Requires 2.1 x 10 mm Universal Sentry Guard Holder, Part No. WAT097958 13 Requires Guard-Pak Holders, Part No. WAT088141

162 www.waters.com/order Nova-Pak Analytical Columns*

— Guard-Pak Inserts 4 µm 10/pk WAT01522013 WAT03588013 WAT02079513 WAT02080013 WAT02079013 WAT08814113 WAT0375256 — 10/pk 4 µm Guard-Pak Inserts — 

 See page 145 for product information. See page 145 for product information. A 4 Requires 3.0 x 20 mm/4.6 x 20 mm Universal Sentry Guard Holder, Part No. WAT046910 C HPLC nalytical 6 Requires Cartridge End-Fittings, Part No. WAT037525 13 Requires Guard-Pak Holder, Part No. WAT088141 * Recommended for Triglycerides and Cholesterol Analysis, see page 156

Shodex RSpak D Series Column Base-Polymer Functional Group Dimension Part No. DS-613 Polystyrene None 6 x 150 mm WAT034220 C and olumns DE-613 Polymethacrylate None 6 x 150 mm WAT034221 DC-613 Polystyrene Sulfonated 6 x 150 mm WAT034223 DS-G Pre-Column 4.6 x 10 mm WAT034224 DE-G Pre-Column 4.6 x 10 mm WAT034225 DC-G Pre-Column 4.6 x 10 mm WAT034227

 See page 149 for product information. onsumables

Standards Standard Neutrals Suitability Standard 186006360

Reversed-Phase Suitability Standard 186006363  See page 310 for product information.

Did you know... Product care and use information is available on our website! Go to: www.waters.com, under "Services and Support" look for "User Guides" or go to www.waters.com/chemcu

163 SunFire Analytical Columns

Dimension Type Particle Size Qty. C18 C8 Dimension Type Particle Size Qty. C18 C8 1.0 x 50 mm Column 2.5 µm 1/pk 186003392 186003394 4.6 x 20 mm Guard 3.5 µm 2/pk 1860026824 1860027274 2.1 x 10 mm Guard 2.5 µm 2/pk 1860033953 1860033963 4.6 x 20 mm IS Column 3.5 µm 1/pk 186002549 186002699 2.1 x 20 mm IS Column 2.5 µm 1/pk 186003397 186003398 4.6 x 30 mm Column 3.5 µm 1/pk 186002550 186002728 2.1 x 30 mm Column 2.5 µm 1/pk 186003399 186003400 4.6 x 50 mm Column 3.5 µm 1/pk 186002551 186002729 2.1 x 50 mm Column 2.5 µm 1/pk 186003401 186003402 4.6 x 75 mm Column 3.5 µm 1/pk 186002552 186002730 2.1 x 75 mm Column 2.5 µm 1/pk 186005634 186005635 4.6 x 100 mm Column 3.5 µm 1/pk 186002553 186002731 3.0 x 20 mm Guard 2.5 µm 2/pk 1860034054 1860034064 4.6 x 150 mm Column 3.5 µm 1/pk 186002554 186002732 3.0 x 20 mm IS Column 2.5 µm 1/pk 186003403 186003404 1.0 x 150 mm Column 5 µm 1/pk 186002529 186002707 3.0 x 30 mm Column 2.5 µm 1/pk 186003407 186003408 2.1 x 10 mm Guard 5 µm 2/pk 1860025363 1860027133 3.0 x 50 mm Column 2.5 µm 1/pk 186003409 186003410 2.1 x 20 mm IS Column 5 µm 1/pk 186002537 186002698 3.0 x 75 mm Column 2.5 µm 1/pk 186005636 186005637 2.1 x 30 mm Column 5 µm 1/pk 186002538 186002714 4.6 x 20 mm Guard 2.5 µm 2/pk 1860034134 1860034144 2.1 x 50 mm Column 5 µm 1/pk 186002539 186002715 4.6 x 20 mm IS Column 2.5 µm 1/pk 186003411 186003412 2.1 x 100 mm Column 5 µm 1/pk 186002540 186002716 4.6 x 30 mm Column 2.5 µm 1/pk 186003415 186003416 2.1 x 150 mm Column 5 µm 1/pk 186002541 186002717 4.6 x 50 mm Column 2.5 µm 1/pk 186003417 186003418 3.0 x 20 mm Guard 5 µm 2/pk 1860026834 1860027224 4.6 x 75 mm Column 2.5 µm 1/pk 186003419 186003420 3.0 x 20 mm IS Column 5 µm 1/pk 186002685 186002702 1.0 x 50 mm Column 3.5 µm 1/pk 186002526 186002705 3.0 x 50 mm Column 5 µm 1/pk 186002545 186002723 1.0 x 150 mm Column 3.5 µm 1/pk 186002528 186002706 3.0 x 100 mm Column 5 µm 1/pk 186002546 186002724 2.1 x 10 mm Guard 3.5 µm 2/pk 1860025303 1860027083 3.0 x 150 mm Column 5 µm 1/pk 186002547 186002725 2.1 x 20 mm IS Column 3.5 µm 1/pk 186002531 186002697 3.0 x 250 mm Column 5 µm 1/pk 186002548 186002726 2.1 x 30 mm Column 3.5 µm 1/pk 186002532 186002709 4.6 x 20 mm Guard 5 µm 2/pk 1860026844 1860027334 onsumables 2.1 x 50 mm Column 3.5 µm 1/pk 186002533 186002710 4.6 x 20 mm IS Column 5 µm 1/pk 186002555 186002700 2.1 x 100 mm Column 3.5 µm 1/pk 186002534 186002711 4.6 x 30 mm Column 5 µm 1/pk 186002556 186002734 2.1 x 150 mm Column 3.5 µm 1/pk 186002535 186002712 4.6 x 50 mm Column 5 µm 1/pk 186002557 186002735 3.0 x 20 mm Guard 3.5 µm 2/pk 1860026814 1860027184 4.6 x 100 mm Column 5 µm 1/pk 186002558 186002736 3.0 x 20 mm IS Column 3.5 µm 1/pk 186002686 186002701 4.6 x 150 mm Column 5 µm 1/pk 186002559 186002737 3.0 x 30 mm Column 3.5 µm 1/pk 186003254 Custom 4.6 x 250 mm Column 5 µm 1/pk 186002560 186002738 olumns and C 3.0 x 50 mm Column 3.5 µm 1/pk 186002542 186002719 3.0 x 100 mm Column 3.5 µm 1/pk 186002543 186002720

3.0 x 150 mm Column 3.5 µm 1/pk 186002544 186002721  See page 136 for product information. 3 Requires 2.1 x 10 mm Universal Sentry Guard Column Holder, Part No. WAT097958 4 Requires 3.0 x 20 mm/4.6 x 20 mm Universal Sentry Guard Holder, Part No. WAT046910 nalytical HPLC C A SunFire Columns Method Validation Kits Each Method Validation Kit contains 3 columns, each from a different batch.

Dimension Particle Size C18 C8 2.1 x 100 mm 3.5 µm 186002674 186002739 4.6 x 100 mm 3.5 µm 186002675 186002740 4.6 x 150 mm 3.5 µm 186002676 186002741 2.1 x 150 mm 5 µm 186002678 186002743 4.6 x 100 mm 5 µm 186002677 186002742 4.6 x 150 mm 5 µm 186002679 186002744

4.6 x 250 mm 5 µm 186002680 186002745  See page 136 for product information.

164 www.waters.com/order A HPLC C HPLC nalytical C and olumns onsumables

Successful Separations Start with Selectivity

XSelect™ HPLC columns are designed for the widest range of chromatographic separations.

■ Two particle technologies (HSS and CSH™)

■ Eight selectivity-optimized chemistries

■ Three scalable particle sizes (2.5, 3.5 and 5 µm)

■ Forty column dimensions providing more than 500 column configurations

Discover what XSelectivity will do for you. www.waters.com/xselect

©2011 Waters Corporation. Waters, XSelect, CSH and The Science of What’s Possible are trademarks of Waters Corporation. 165 Symmetry, SymmetryShield, and Symmetry300 Analytical Columns Particle Dimension Type Qty. Symmetry C Symmetry C SymmetryShield RP18 SymmetryShield RP8 Symmetry300 C Symmetry300 C Size 18 8 18 4 1.0 x 50 mm Column 3.5 µm 1/pk WAT106056 WAT106052 186000175 WAT106060 186000185 186000276 1.0 x 150 mm Column 3.5 µm 1/pk WAT248059 WAT248072 186000176 WAT106048 — — 2.1 x 10 mm Guard 3.5 µm 2/pk WAT1061273 WAT1061283 1860001693 WAT1061293 1860001983 1860002753 2.1 x 20 mm Cartridge 3.5 µm 1/pk 18600026911 18600026811 — — — — 2.1 x 20 mm IS Column 3.5 µm 1/pk 186002066 186002067 186002068 186002069 — — 2.1 x 30 mm Column 3.5 µm 1/pk WAT058973 WAT058977 186000171 WAT106001 — — 2.1 x 50 mm Cartridge 3.5 µm 1/pk 18600015211 18600014911 18600016811 18600014711 18600019911 — 2.1 x 50 mm Column 3.5 µm 1/pk WAT200650 WAT200624 186000172 WAT094257 186000187 186000277 2.1 x 100 mm Cartridge 3.5 µm 1/pk 18600015111 18600015311 18600016711 18600014611 — — 2.1 x 100 mm Column 3.5 µm 1/pk WAT058965 WAT058961 186000173 WAT058969 186000188 186000278 2.1 x 150 mm Cartridge 3.5 µm 1/pk 18600015011 18600014811 18600016611 18600014511 — — 2.1 x 150 mm Column 3.5 µm 1/pk WAT106005 WAT106011 186000174 WAT106008 186000200 186000279 3.0 x 20 mm IS Column 3.5 µm 1/pk 186002074 186002075 186002076 186002077 — — 3.0 x 50 mm Column 3.5 µm 1/pk 186002612 186002613 186002614 186002615 — — 3.0 x 100 mm Column 3.5 µm 1/pk 186000696 186000698 186000700 186000703 — — 3.0 x 150 mm Column 3.5 µm 1/pk 186000695 186000697 186000699 186000702 — — 3.9 x 20 mm Guard 3.5 µm 2/pk — — 18600070112 18600070412 — — 3.9 x 20 mm IS Column 3.5 µm 1/pk 186002082 186002083 186002084 186002085 — — 4.6 x 20 mm IS Column 3.5 µm 1/pk 186002090 186002091 186002092 186002093 — — 4.6 x 30 mm Column 3.5 µm 1/pk 186000271 186000270 — — — — 4.6 x 50 mm Column 3.5 µm 1/pk WAT200625 WAT200620 186000177 WAT094260 186000201 186000280 4.6 x 75 mm Cartridge 3.5 µm 1/pk WAT06626011 WAT06621011 18600018311 WAT09427211 18600019211 — onsumables 4.6 x 75 mm Column 3.5 µm 1/pk WAT066224 WAT066200 186000178 WAT094263 186000189 186000281 4.6 x 100 mm Cartridge 3.5 µm 1/pk WAT06626511 WAT06621511 18600017011 WAT09427511 — — 4.6 x 100 mm Column 3.5 µm 1/pk WAT066220 WAT066204 186000179 WAT094266 186000190 186000282 4.6 x 150 mm Column 3.5 µm 1/pk WAT200632 WAT200630 186000180 WAT094269 186000197 186000283 4.6 x 250 mm Column 3.5 µm 1/pk 186005794 2.1 x 20 mm IS Column 5 µm 1/pk 186002070 186002071 186002072 186002073 — — olumns and C 2.1 x 50 mm Column 5 µm 1/pk 186000206 186000212 186000217 186000223 — — 2.1 x 100 mm Column 5 µm 1/pk 186002608 186002609 186000998 186002611 — — 2.1 x 150 mm Column 5 µm 1/pk WAT056975 WAT056955 186000111 WAT094245 WAT106172 186000285 3.0 x 20 mm IS Column 5 µm 1/pk 186002078 186002079 186002080 186002081 — — 3.0 x 150 mm Column 5 µm 1/pk WAT054200 WAT054230 186000692 WAT094243 — — 3.0 x 250 mm Column 5 µm 1/pk 186000690 186000691 186000693 186000694 — — 3.9 x 20 mm Guard 5 µm 2/pk WAT05422512 WAT05425012 18600010712 WAT20067512 WAT10616612 18600028412

11 11 11

nalytical HPLC C 3.9 x 50 mm Cartridge 5 µm 1/pk WAT054220 WAT054240 — WAT094248 — —

A 3.9 x 20 mm IS Column 5 µm 1/pk 186002086 186002087 186002088 186002089 — — 3.9 x 150 mm Cartridge 5 µm 1/pk WAT05420511 WAT05423511 18600010611 WAT20065811 WAT10616911 — 3.9 x 150 mm Column 5 µm 1/pk WAT046980 WAT046970 186000108 WAT200655 WAT106154 186000286 4.6 x 20 mm IS Column 5 µm 1/pk 186002094 186002095 186002096 186002097 — — 4.6 x 50 mm Column 5 µm 1/pk 186000207 186000213 186000218 186000224 WAT106209 186000287 4.6 x 100 mm Column 5 µm 1/pk 186002616 186002617 186002618 186002619 — — 4.6 x 150 mm Cartridge 5 µm 1/pk WAT05421011 WAT05425511 18600011011 WAT20066511 WAT10616311 — 4.6 x 150 mm Column 5 µm 1/pk WAT045905 WAT045995 186000109 WAT200662 WAT106157 186000288 4.6 x 250 mm Cartridge 5 µm 1/pk WAT05421511 WAT05424511 18600011311 WAT20066111 WAT10616011 —

4.6 x 250 mm Column 5 µm 2/pk WAT054275 WAT054270 186000112 WAT200670 WAT106151 186000289  See page 141 for product information. 3 Requires 2.1 x 10 mm Sentry Guard Holder, Part No. WAT097958 11 Requires Cartridge End-Fittings, see page 389 12 Requires Sentry Guard Holders, see page 148

166 www.waters.com/order Symmetry NanoEase Nano and Capillary Columns

Length Type Column ID Particle Size Qty. Symmetry C18 Symmetry300 C18 50 mm Column 75 µm 3.5 µm 1/pk 186002188 186002191 100 mm Column 75 µm 3.5 µm 1/pk 186002189 186002192 150 mm Column 75 µm 3.5 µm 1/pk 186002190 186002193 50 mm Column 100 µm 3.5 µm 1/pk 186002201 — 100 mm Column 100 µm 3.5 µm 1/pk 186002202 — 150 mm Column 100 µm 3.5 µm 1/pk 186002203 186002206 50 mm Column 150 µm 3.5 µm 1/pk 186002459 186002462 100 mm Column 150 µm 3.5 µm 1/pk 186002461 186002463 150 mm Column 150 µm 3.5 µm 1/pk — 186002464 50 mm Column 300 µm 3.5 µm 1/pk 186002581 186002587 100 mm Column 300 µm 3.5 µm 1/pk 186002582 186002588 150 mm Column 300 µm 3.5 µm 1/pk 186002583 186002589 50 mm Column 300 µm 5 µm 1/pk 186002584 186002590 100 mm Column 300 µm 5 µm 1/pk 186002585 186002591

150 mm Column 300 µm 5 µm 1/pk 186002586 186002592  See page 141 for product information. A HPLC C HPLC nalytical

Symmetry, SymmetryShield, and Symmetry300 Method Validation Kits Each Method Validation Kit contains 3 columns, each from a different batch.

Dimension Type Particle Size Symmetry C18 Symmetry C8 SymmetryShield RP18 SymmetryShield RP8 Symmetry300 C18 Symmetry300 C4 2.1 x 150 mm Column 3.5 µm — — 186000182 — 186000194 186000290 4.6 x 150 mm Column 3.5 µm WAT094240 WAT094237 186000181 WAT094278 186000195 186000291 2.1 x 150 mm Column 5 µm WAT094234 WAT094231 186000100 WAT094254 WAT106193 186000292

3.0 x 150 mm Column 5 µm WAT054446 WAT054434 — WAT094251 — — C and olumns 3.9 x 150 mm Cartridge 5 µm WAT05445211 WAT05444011 18600010511 WAT21058211 WAT10618111 — 3.9 x 150 mm Column 5 µm WAT047210 WAT046955 186000104 WAT210594 WAT106187 186000293 4.6 x 150 mm Cartridge 5 µm WAT05445411 WAT05444211 18600010111 WAT21058511 WAT10617511 — 4.6 x 150 mm Column 5 µm WAT054448 WAT054435 186000103 WAT210588 WAT106190 186000294 4.6 x 250 mm Cartridge 5 µm WAT05445611 WAT05444411 18600011411 WAT21057911 WAT10617811 —

4.6 x 250 mm Column 5 µm WAT054450 WAT054438 186000102 WAT210591 WAT106184 186000295 onsumables  See page 141 for product information. 11 Requires Cartridge End-Fittings, see page 389

VanGuard Pre-Columns(Guard Columns) Part No. Chemistry Particle Size Dimension 3/pk

XBridge BEH C18 2.5 μm 2.1 x 5 mm 186006291 XBridge BEH Shield RP18 2.5 μm 2.1 x 5 mm 186006293

XBridge BEH C8 2.5 μm 2.1 x 5 mm 186006292 XBridge BEH Phenyl 2.5 μm 2.1 x 5 mm 186006294 XBridge BEH HILIC 2.5 μm 2.1 x 5 mm 186006295 XBridge BEH Amide 2.5 μm 2.1 x 5 mm 186006296

XSelect CSH C18 2.5 μm 2.1 x 5 mm 186006297 XSelect CSH Fluoro-Phenyl 2.5 μm 2.1 x 5 mm 186006298 XSelect CSH Phenyl-Hexyl 2.5 μm 2.1 x 5 mm 186006299 XSelect HSS T3 2.5 μm 2.1 x 5 mm 186006301

XSelect HSS C18 2.5 μm 2.1 x 5 mm 186006300

XSelect HSS C18 SB 2.5 μm 2.1 x 5 mm 186006302 XSelect HSS Cyano 2.5 μm 2.1 x 5 mm 186006304

XSelect HSS PFP 2.5 μm 2.1 x 5 mm 186006303  See page 103 for product information.

167 Waters Spherisorb Columns

Dimension Type Particle Size Qty. ODS1 ODS2 ODS B C8 C6 C1 NH2 Phenyl CN Normal Phase CN Reversed Phase W Silica SAX SCX OD/CN Qty. Particle Size Type Dimension 1 x 100 mm Column 3 µm 1/pk — PSS832132 — PSS832232 — — — — PSS832432 — PSS832032 — — — 1/pk 3 µm Column 1 x 100 mm 1 x 150 mm Column 3 µm 1/pk PSS833433 PSS832133 — PSS832233 — — PSS832333 — PSS832433 — PSS832033 — — — 1/pk 3 µm Column 1 x 150 mm 2 x 100 mm Column 3 µm 1/pk PSS833422 PSS832122 — PSS832222 — — PSS832322 — PSS832422 — PSS832022 — — — 1/pk 3 µm Column 2 x 100 mm 2 x 150 mm Column 3 µm 1/pk PSS833423 PSS832123 — PSS832223 — — PSS832323 — PSS832423 — PSS832023 — — — 1/pk 3 µm Column 2 x 150 mm 4.6 x 30 mm Cartridge 3 µm 1/pk — PSS8300656 — — — — — — — — PSS8300686 — — — 1/pk 3 µm Cartridge 4.6 x 30 mm 4.6 x 50 mm Cartridge 3 µm 1/pk — PSS8395466 — PSS8395516 PSS8395816 PSS8395766 PSS8395566 PSS8395916 PSS8395616 — PSS8395416 — — — 1/pk 3 µm Cartridge 4.6 x 50 mm 4.6 x 50 mm Column 3 µm 1/pk PSS833411 PSS832111 — PSS832211 PSS833111 PSS832911 PSS832311 PSS833711 PSS832411 PSS832011 — — — 1/pk 3 µm Column 4.6 x 50 mm 4.6 x 60 mm Column 3 µm 1/pk PSS839852 PSS839853 — — — — — — — — — — — — 1/pk 3 µm Column 4.6 x 60 mm 4.6 x 100 mm Cartridge 3 µm 1/pk PSS8395876 PSS8395476 — PSS8395526 PSS8395826 PSS8395776 PSS8395576 PSS8395926 PSS8395626 — PSS8395426 — — — 1/pk 3 µm Cartridge 4.6 x 100 mm 4.6 x 100 mm Column 3 µm 1/pk PSS833412 PSS832112 — PSS832212 PSS833112 PSS832912 PSS832312 PSS833712 PSS832412 — PSS832012 — — — 1/pk 3 µm Column 4.6 x 100 mm 4.6 x 150 mm Cartridge 3 µm 1/pk PSS8395886 PSS8395486 — PSS8395536 PSS8395836 PSS8395786 PSS8395586 PSS8395936 PSS8395636 — PSS8395436 — — — 1/pk 3 µm Cartridge 4.6 x 150 mm 4.6 x 150 mm Column 3 µm 1/pk PSS833413 PSS832113 — PSS832213 PSS833113 PSS832913 PSS832313 PSS833713 PSS832413 — PSS832013 — — — 1/pk 3 µm Column 4.6 x 150 mm 1 x 100 mm Column 5 µm 1/pk PSS830632 PSS831932 — PSS831832 PSS831032 — PSS831132 — PSS830932 — — — — — 1/pk 5 µm Column 1 x 100 mm 1 x 150 mm Column 5 µm 1/pk — PSS831933 — — — — — — — — — — — — 1/pk 5 µm Column 1 x 150 mm 1 x 250 mm Column 5 µm 1/pk PSS830635 PSS831935 — — — — PSS831135 — — — PSS830135 — — — 1/pk 5 µm Column 1 x 250 mm 2 x 100 mm Column 5 µm 1/pk PSS830622 PSS831922 — PSS831822 — — PSS831122 — PSS830922 — PSS830122 — — — 1/pk 5 µm Column 2 x 100 mm 2 x 150 mm Column 5 µm 1/pk PSS830623 PSS831923 — PSS831823 — — PSS831123 — PSS830923 — PSS830123 — — — 1/pk 5 µm Column 2 x 150 mm 2 x 250 mm Column 5 µm 1/pk PSS830625 PSS831925 — PSS831825 — — PSS831125 — PSS830925 — PSS830125 — — — 1/pk 5 µm Column 2 x 250 mm 10 x 4.6 mm Guard 5 µm 3/pk PSS8300738 PSS8300538 PSS8300598 PSS8300748 PSS8300758 PSS8300768 PSS8300798 PSS8300788 PSS8300778 — PSS8300518 PSS8300558 PSS8300578 — 3/pk 5 µm Guard 10 x 4.6 mm 30 x 4.6 mm Guard 5 µm 3/pk PSS8394728 PSS8394588 PSS8394708 PSS8394738 PSS8394748 PSS8394758 PSS8394788 PSS8394778 PSS8394768 — PSS8394518 PSS8394658 PSS8394718 — 3/pk 5 µm Guard 30 x 4.6 mm 3.0 x 125 mm Cartridge 5 µm 1/pk PSS8385306 PSS8385296 — PSS8385336 — — PSS8385356 — PSS8385316 — PSS8385346 — — — 1/pk 5 µm Cartridge 3.0 x 125 mm onsumables 3.0 x 250 mm Cartridge 5 µm 1/pk PSS8385236 PSS8385226 — PSS8385256 — — PSS8385266 — PSS8385246 — PSS8385216 186006836 — — 1/pk 5 µm Cartridge 3.0 x 250 mm 4.0 x 125 mm Cartridge 5 µm 1/pk PSS8455516 PSS8455536 — PSS8455556 PSS8455576 PSS8455596 PSS8455656 PSS8455616 PSS8455636 — PSS8455496 PSS8455676 PSS8455696 — 1/pk 5 µm Cartridge 4.0 x 125 mm 4.0 x 125 mm Column 5 µm 1/pk PSS845541 PSS845543 — PSS845280 PSS845284 PSS845288 PSS845300 PSS845292 PSS845296 — PSS845539 PSS845304 PSS845308 — 1/pk 5 µm Column 4.0 x 125 mm 4.0 x 250 mm Cartridge 5 µm 1/pk PSS8455526 PSS8455546 — PSS8455566 PSS8455586 PSS8455606 PSS8455666 PSS8455626 PSS8455646 — PSS8455506 PSS8455686 PSS8455706 — 1/pk 5 µm Cartridge 4.0 x 250 mm 4.0 x 250 mm Column 5 µm 1/pk PSS845542 PSS845277 — PSS845281 PSS845285 PSS845289 PSS845301 PSS845293 PSS845297 — PSS845540 PSS845305 PSS845309 — 1/pk 5 µm Column 4.0 x 250 mm 4.6 x 50 mm Cartridge 5 µm 1/pk PSS8395066 PSS8395366 — PSS8395316 PSS8395216 PSS8395666 PSS8395266 PSS8395116 PSS8395166 — — PSS8395716 PSS8396516 — 1/pk 5 µm Cartridge 4.6 x 50 mm olumns and C 4.6 x 50 mm Column 5 µm 1/pk PSS830611 PSS831911 — PSS831811 PSS831011 PSS832611 PSS831111 — PSS830911 — PSS830111 PSS832711 PSS837511 — 1/pk 5 µm Column 4.6 x 50 mm 4.6 x 100 mm Cartridge 5 µm 1/pk PSS8395076 PSS8395376 — PSS8395326 PSS8395226 PSS8395676 PSS8395276 PSS8395126 PSS8395176 — PSS8395026 PSS8395726 PSS8396526 — 1/pk 5 µm Cartridge 4.6 x 100 mm 4.6 x 100 mm Column 5 µm 1/pk PSS830612 PSS831912 — PSS831812 PSS831012 PSS832612 PSS831112 PSS830812 PSS830912 — PSS830112 PSS832712 PSS837512 PSS837812 1/pk 5 µm Column 4.6 x 100 mm 4.6 x 150 mm Cartridge 5 µm 1/pk PSS8395086 PSS8395386 PSS8396136 PSS8395336 PSS8395236 PSS8395686 PSS8395286 PSS8395136 PSS8395186 — PSS8395036 PSS8395736 PSS8396536 PSS8396436 1/pk 5 µm Cartridge 4.6 x 150 mm 4.6 x 150 mm Column 5 µm 1/pk PSS830613 PSS831913 — PSS831813 PSS831013 PSS832613 PSS831113 PSS830813 PSS830913 PSS830908 PSS830113 PSS832713 PSS837513 PSS837813 1/pk 5 µm Column 4.6 x 150 mm 4.6 x 250 mm Cartridge 5 µm 1/pk PSS8395106 PSS8395406 PSS8396156 PSS8395356 PSS8395256 PSS8395706 PSS8395306 PSS8395156 PSS8395206 — PSS8395056 PSS8395756 PSS8396556 PSS8396456 1/pk 5 µm Cartridge 4.6 x 250 mm 4.6 x 250 mm Column 5 µm 1/pk PSS830615 PSS831915 — PSS831815 PSS831015 PSS832615 PSS831115 PSS830815 PSS830915 PSS830909 PSS830115 PSS832715 PSS837515 PSS837815 1/pk 5 µm Column 4.6 x 250 mm

nalytical HPLC C 4.6 x 100 mm Column 10 µm 1/pk PSS830712 — — — — — PSS833612 — PSS833512 — PSS830212 PSS833912 — — 1/pk 10 µm Column 4.6 x 100 mm

A 4.6 x 150 mm Column 10 µm 1/pk PSS830713 PSS832513 — PSS832813 PSS833213 PSS833013 PSS833613 PSS833813 PSS833513 — PSS830213 PSS833913 PSS837613 — 1/pk 10 µm Column 4.6 x 150 mm 4.6 x 200 mm Column 10 µm 1/pk PSS830714 PSS832514 — — — — — — PSS832514 — PSS830214 — — — 1/pk 10 µm Column 4.6 x 200 mm

4.6 x 250 mm Column 10 µm 1/pk PSS830715 PSS832515 — PSS832815 PSS833215 PSS833015 PSS833615 PSS833815 PSS833515 — PSS830215 PSS833915 PSS837615 — 1/pk 10 µm Column 4.6 x 250 mm

  See page 144 for product information. See page 144 for product information. 6 Requires End Connector Kit, Part No. PSS614100 8 Requires Inline Guard Cartridge Holder, Part No. PSS830008

168 www.waters.com/order Waters Spherisorb Columns

Dimension Type Particle Size Qty. ODS1 ODS2 ODS B C8 C6 C1 NH2 Phenyl CN Normal Phase CN Reversed Phase W Silica SAX SCX OD/CN Qty. Particle Size Type Dimension 1 x 100 mm Column 3 µm 1/pk — PSS832132 — PSS832232 — — — — PSS832432 — PSS832032 — — — 1/pk 3 µm Column 1 x 100 mm 1 x 150 mm Column 3 µm 1/pk PSS833433 PSS832133 — PSS832233 — — PSS832333 — PSS832433 — PSS832033 — — — 1/pk 3 µm Column 1 x 150 mm 2 x 100 mm Column 3 µm 1/pk PSS833422 PSS832122 — PSS832222 — — PSS832322 — PSS832422 — PSS832022 — — — 1/pk 3 µm Column 2 x 100 mm 2 x 150 mm Column 3 µm 1/pk PSS833423 PSS832123 — PSS832223 — — PSS832323 — PSS832423 — PSS832023 — — — 1/pk 3 µm Column 2 x 150 mm 4.6 x 30 mm Cartridge 3 µm 1/pk — PSS8300656 — — — — — — — — PSS8300686 — — — 1/pk 3 µm Cartridge 4.6 x 30 mm 4.6 x 50 mm Cartridge 3 µm 1/pk — PSS8395466 — PSS8395516 PSS8395816 PSS8395766 PSS8395566 PSS8395916 PSS8395616 — PSS8395416 — — — 1/pk 3 µm Cartridge 4.6 x 50 mm 4.6 x 50 mm Column 3 µm 1/pk PSS833411 PSS832111 — PSS832211 PSS833111 PSS832911 PSS832311 PSS833711 PSS832411 PSS832011 — — — 1/pk 3 µm Column 4.6 x 50 mm 4.6 x 60 mm Column 3 µm 1/pk PSS839852 PSS839853 — — — — — — — — — — — — 1/pk 3 µm Column 4.6 x 60 mm 4.6 x 100 mm Cartridge 3 µm 1/pk PSS8395876 PSS8395476 — PSS8395526 PSS8395826 PSS8395776 PSS8395576 PSS8395926 PSS8395626 — PSS8395426 — — — 1/pk 3 µm Cartridge 4.6 x 100 mm 4.6 x 100 mm Column 3 µm 1/pk PSS833412 PSS832112 — PSS832212 PSS833112 PSS832912 PSS832312 PSS833712 PSS832412 — PSS832012 — — — 1/pk 3 µm Column 4.6 x 100 mm 4.6 x 150 mm Cartridge 3 µm 1/pk PSS8395886 PSS8395486 — PSS8395536 PSS8395836 PSS8395786 PSS8395586 PSS8395936 PSS8395636 — PSS8395436 — — — 1/pk 3 µm Cartridge 4.6 x 150 mm 4.6 x 150 mm Column 3 µm 1/pk PSS833413 PSS832113 — PSS832213 PSS833113 PSS832913 PSS832313 PSS833713 PSS832413 — PSS832013 — — — 1/pk 3 µm Column 4.6 x 150 mm 1 x 100 mm Column 5 µm 1/pk PSS830632 PSS831932 — PSS831832 PSS831032 — PSS831132 — PSS830932 — — — — — 1/pk 5 µm Column 1 x 100 mm 1 x 150 mm Column 5 µm 1/pk — PSS831933 — — — — — — — — — — — — 1/pk 5 µm Column 1 x 150 mm 1 x 250 mm Column 5 µm 1/pk PSS830635 PSS831935 — — — — PSS831135 — — — PSS830135 — — — 1/pk 5 µm Column 1 x 250 mm 2 x 100 mm Column 5 µm 1/pk PSS830622 PSS831922 — PSS831822 — — PSS831122 — PSS830922 — PSS830122 — — — 1/pk 5 µm Column 2 x 100 mm A 2 x 150 mm Column 5 µm 1/pk PSS830623 PSS831923 — PSS831823 — — PSS831123 — PSS830923 — PSS830123 — — — 1/pk 5 µm Column 2 x 150 mm HPLC C HPLC nalytical 2 x 250 mm Column 5 µm 1/pk PSS830625 PSS831925 — PSS831825 — — PSS831125 — PSS830925 — PSS830125 — — — 1/pk 5 µm Column 2 x 250 mm 10 x 4.6 mm Guard 5 µm 3/pk PSS8300738 PSS8300538 PSS8300598 PSS8300748 PSS8300758 PSS8300768 PSS8300798 PSS8300788 PSS8300778 — PSS8300518 PSS8300558 PSS8300578 — 3/pk 5 µm Guard 10 x 4.6 mm 30 x 4.6 mm Guard 5 µm 3/pk PSS8394728 PSS8394588 PSS8394708 PSS8394738 PSS8394748 PSS8394758 PSS8394788 PSS8394778 PSS8394768 — PSS8394518 PSS8394658 PSS8394718 — 3/pk 5 µm Guard 30 x 4.6 mm 3.0 x 125 mm Cartridge 5 µm 1/pk PSS8385306 PSS8385296 — PSS8385336 — — PSS8385356 — PSS8385316 — PSS8385346 — — — 1/pk 5 µm Cartridge 3.0 x 125 mm 3.0 x 250 mm Cartridge 5 µm 1/pk PSS8385236 PSS8385226 — PSS8385256 — — PSS8385266 — PSS8385246 — PSS8385216 186006836 — — 1/pk 5 µm Cartridge 3.0 x 250 mm 4.0 x 125 mm Cartridge 5 µm 1/pk PSS8455516 PSS8455536 — PSS8455556 PSS8455576 PSS8455596 PSS8455656 PSS8455616 PSS8455636 — PSS8455496 PSS8455676 PSS8455696 — 1/pk 5 µm Cartridge 4.0 x 125 mm 4.0 x 125 mm Column 5 µm 1/pk PSS845541 PSS845543 — PSS845280 PSS845284 PSS845288 PSS845300 PSS845292 PSS845296 — PSS845539 PSS845304 PSS845308 — 1/pk 5 µm Column 4.0 x 125 mm C and olumns 4.0 x 250 mm Cartridge 5 µm 1/pk PSS8455526 PSS8455546 — PSS8455566 PSS8455586 PSS8455606 PSS8455666 PSS8455626 PSS8455646 — PSS8455506 PSS8455686 PSS8455706 — 1/pk 5 µm Cartridge 4.0 x 250 mm 4.0 x 250 mm Column 5 µm 1/pk PSS845542 PSS845277 — PSS845281 PSS845285 PSS845289 PSS845301 PSS845293 PSS845297 — PSS845540 PSS845305 PSS845309 — 1/pk 5 µm Column 4.0 x 250 mm 4.6 x 50 mm Cartridge 5 µm 1/pk PSS8395066 PSS8395366 — PSS8395316 PSS8395216 PSS8395666 PSS8395266 PSS8395116 PSS8395166 — — PSS8395716 PSS8396516 — 1/pk 5 µm Cartridge 4.6 x 50 mm 4.6 x 50 mm Column 5 µm 1/pk PSS830611 PSS831911 — PSS831811 PSS831011 PSS832611 PSS831111 — PSS830911 — PSS830111 PSS832711 PSS837511 — 1/pk 5 µm Column 4.6 x 50 mm 4.6 x 100 mm Cartridge 5 µm 1/pk PSS8395076 PSS8395376 — PSS8395326 PSS8395226 PSS8395676 PSS8395276 PSS8395126 PSS8395176 — PSS8395026 PSS8395726 PSS8396526 — 1/pk 5 µm Cartridge 4.6 x 100 mm 4.6 x 100 mm Column 5 µm 1/pk PSS830612 PSS831912 — PSS831812 PSS831012 PSS832612 PSS831112 PSS830812 PSS830912 — PSS830112 PSS832712 PSS837512 PSS837812 1/pk 5 µm Column 4.6 x 100 mm onsumables 4.6 x 150 mm Cartridge 5 µm 1/pk PSS8395086 PSS8395386 PSS8396136 PSS8395336 PSS8395236 PSS8395686 PSS8395286 PSS8395136 PSS8395186 — PSS8395036 PSS8395736 PSS8396536 PSS8396436 1/pk 5 µm Cartridge 4.6 x 150 mm 4.6 x 150 mm Column 5 µm 1/pk PSS830613 PSS831913 — PSS831813 PSS831013 PSS832613 PSS831113 PSS830813 PSS830913 PSS830908 PSS830113 PSS832713 PSS837513 PSS837813 1/pk 5 µm Column 4.6 x 150 mm 4.6 x 250 mm Cartridge 5 µm 1/pk PSS8395106 PSS8395406 PSS8396156 PSS8395356 PSS8395256 PSS8395706 PSS8395306 PSS8395156 PSS8395206 — PSS8395056 PSS8395756 PSS8396556 PSS8396456 1/pk 5 µm Cartridge 4.6 x 250 mm 4.6 x 250 mm Column 5 µm 1/pk PSS830615 PSS831915 — PSS831815 PSS831015 PSS832615 PSS831115 PSS830815 PSS830915 PSS830909 PSS830115 PSS832715 PSS837515 PSS837815 1/pk 5 µm Column 4.6 x 250 mm 4.6 x 100 mm Column 10 µm 1/pk PSS830712 — — — — — PSS833612 — PSS833512 — PSS830212 PSS833912 — — 1/pk 10 µm Column 4.6 x 100 mm 4.6 x 150 mm Column 10 µm 1/pk PSS830713 PSS832513 — PSS832813 PSS833213 PSS833013 PSS833613 PSS833813 PSS833513 — PSS830213 PSS833913 PSS837613 — 1/pk 10 µm Column 4.6 x 150 mm 4.6 x 200 mm Column 10 µm 1/pk PSS830714 PSS832514 — — — — — — PSS832514 — PSS830214 — — — 1/pk 10 µm Column 4.6 x 200 mm

4.6 x 250 mm Column 10 µm 1/pk PSS830715 PSS832515 — PSS832815 PSS833215 PSS833015 PSS833615 PSS833815 PSS833515 — PSS830215 PSS833915 PSS837615 — 1/pk 10 µm Column 4.6 x 250 mm

  See page 144 for product information. See page 144 for product information. 6 Requires End Connector Kit, Part No. PSS614100 8 Requires Inline Guard Cartridge Holder, Part No. PSS830008

169 XBridge Analytical Columns

Dimension Type Particle Size Qty. C18 C8 Shield RP18 Phenyl HILIC Amide 1.0 x 50 mm Column 2.5 µm 1/pk 186003118 186003164 186003136 186003306 — — 2.1 x 10 mm Guard 2.5 µm 2/pk 1860030563 1860030743 1860030653 1860033593 1860044553 — 2.1 x 20 mm IS Column 2.5 µm 1/pk 186003201 186003167 186003139 186003307 — — 2.1 x 30 mm Column 2.5 µm 1/pk 186003084 186003099 186003091 186003308 186004456 — 2.1 x 30 mm XP Column 2.5 μm 1/pk 186006028 186006040 186006052 186006064 186006076 186006088 2.1 x 30 mm XP Column 2.5 μm 3/pk 176002546 176002554 176002562 176002570 176002578 176002586 2.1 x 50 mm Column 2.5 µm 1/pk 186003085 186003101 186003092 186003309 186004457 — 2.1 x 50 mm XP Column 2.5 µm 1/pk 186006029 186006041 186006053 186006065 186006077 186006089 2.1 x 50 mm XP Column 2.5 µm 3/pk 176002547 176002555 176002563 176002571 176002579 176002587 2.1 x 75 mm Column 2.5 µm 1/pk 186005626 186005627 186005628 186005629 — — 2.1 x 75 mm XP Column 2.5 µm 1/pk 186006030 186006042 186006054 186006066 186006078 186006090 2.1 x 75 mm XP Column 2.5 µm 3/pk 176002548 176002556 176002564 176002572 176002580 176002588 2.1 x 100 mm XP Column 2.5 µm 1/pk 186006031 186006043 186006055 186006067 186006079 186006091 2.1 x 100 mm XP Column 2.5 µm 3/pk 176002549 176002557 176002565 176002573 176002581 176002589 2.1 x 150 mm XP Column 2.5 µm 1/pk 186006709 186006712 186006715 186006718 186006721 186006724 2.1 x 150 mm XP Column 2.5 µm 3/pk 176002879 176002881 176002883 176002885 176002887 176002889 3.0 x 20 mm Guard 2.5 µm 2/pk 1860030574 1860030754 1860030664 1860033604 — — 3.0 x 20 mm IS Column 2.5 µm 1/pk 186003087 186003168 186003140 186003310 — — 3.0 x 30 mm Column 2.5 µm 1/pk 186003121 186003169 186003141 186003311 — — 3.0 x 30 mm XP Column 2.5 µm 1/pk 186006032 186006044 186006056 186006068 186006080 186006092 3.0 x 30 mm XP Column 2.5 µm 3/pk 176002550 176002558 176002566 176002574 176002582 176002590 onsumables 3.0 x 50 mm Column 2.5 µm 1/pk 186003122 186003170 186003142 186003312 186004458 — 3.0 x 50 mm XP Column 2.5 µm 1/pk 186006033 186006045 186006057 186006069 186006081 186006093 3.0 x 50 mm XP Column 2.5 µm 3/pk 176002551 176002559 176002567 176002575 176002583 176002591 3.0 x 75 mm Column 2.5 µm 1/pk 186005630 186005631 186005632 186005633 — — 3.0 x 75 mm XP Column 2.5 µm 1/pk 186006034 186006046 186006058 186006070 186006082 186006094

olumns and C 3.0 x 75 mm XP Column 2.5 µm 3/pk 176002552 176002560 176002568 176002576 176002584 176002592 3.0 x 100 mm XP Column 2.5 µm 1/pk 186006035 186006047 186006059 186006071 186006083 186006095 3.0 x 100 mm XP Column 2.5 µm 3/pk 176002553 176002561 176002569 176002577 176002585 176002593 3.0 x 150 mm XP Column 2.5 µm 1/pk 186006710 186006713 186006716 186006719 186006722 186006725 3.0 x 150 mm XP Column 2.5 µm 3/pk 176002880 176002882 176002884 176002886 176002888 176002890 4.6 x 20 mm Guard 2.5 µm 2/pk 1860030584 1860030764 1860030674 1860033614 1860044594 — 4.6 x 20 mm IS Column 2.5 µm 1/pk 186003088 186003172 186003144 186003313 — —

nalytical HPLC C 4.6 x 30 mm Column 2.5 µm 1/pk 186003089 186003173 186003145 186003314 — — A 4.6 x 30 mm XP Column 2.5 µm 1/pk 186006036 186006048 186006060 186006072 186006084 186006096 4.6 x 30 mm XP Column 2.5 µm 3/pk — — — — — — 4.6 x 50 mm Column 2.5 µm 1/pk 186003090 186003174 186003096 186003315 186004460 — 4.6 x 50 mm XP Column 2.5 µm 1/pk 186006037 186006049 186006061 186006073 186006085 186006097 4.6 x 50 mm XP Column 2.5 µm 3/pk — — — — — — 4.6 x 75 mm Column 2.5 µm 1/pk 186003124 186003175 186003146 186003316 186004461 — 4.6 x 75 mm XP Column 2.5 µm 1/pk 186006038 186006050 186006062 186006074 186006086 186006098 4.6 x 75 mm XP Column 2.5 µm 3/pk — — — — — — 4.6 x 100 mm XP Column 2.5 µm 1/pk 186006039 186006051 186006063 186006075 186006087 186006099 4.6 x 100 mm XP Column 2.5 µm 3/pk — — — — — —

4.6 x 150 mm XP Column 2.5 µm 1/pk 186006711 186006714 186006717 186006720 186006723 186006726  See page 132 for product information.

170 www.waters.com/order Dimension Type Particle Size Qty. C18 C8 Shield RP18 Phenyl HILIC Amide 1.0 x 50 mm Column 3.5 µm 1/pk 186003126 186003177 186003148 186003317 186004429 186004871 1.0 x 100 mm Column 3.5 µm 1/pk 186003127 186003178 186003149 186003318 — — 1.0 x 150 mm Column 3.5 µm 1/pk 186003128 186003179 186003150 186003319 — — 2.1 x 10 mm Guard 3.5 µm 2/pk 1860030593 1860030773 1860030683 1860033623 1860044303 1860048573 2.1 x 20 mm IS Column 3.5 µm 1/pk 186003019 186003180 186003151 186003320 — — 2.1 x 30 mm Column 3.5 µm 1/pk 186003020 186003046 186003035 186003321 186004431 186004858 2.1 x 50 mm Column 3.5 µm 1/pk 186003021 186003047 186003036 186003322 186004432 186004859 2.1 x 100 mm Column 3.5 µm 1/pk 186003022 186003048 186003037 186003323 186004433 186004860 2.1 x 150 mm Column 3.5 µm 1/pk 186003023 186003049 186003038 186003324 186004434 186004861 3.0 x 20 mm Guard 3.5 µm 2/pk 1860030604 1860030784 1860030694 1860033634 — — 3.0 x 20 mm IS Column 3.5 µm 1/pk 186003024 186003181 186003152 186003325 — — 3.0 x 30 mm Column 3.5 µm 1/pk 186003025 186003182 186003153 186003326 — 186004862 3.0 x 50 mm Column 3.5 µm 1/pk 186003026 186003050 186003039 186003327 186004435 186004863 3.0 x 100 mm Column 3.5 µm 1/pk 186003027 186003051 186003040 186003328 186004436 186004864 3.0 x 150 mm Column 3.5 µm 1/pk 186003028 186003052 186003041 186003329 — —

4 4 4 4 4 4

4.6 x 20 mm Guard 3.5 µm 2/pk 186003061 186003079 186003070 186003364 186004437 186004865 A

4.6 x 20 mm IS Column 3.5 µm 1/pk 186003029 186003183 186003154 186003330 — — C HPLC nalytical 4.6 x 30 mm Column 3.5 µm 1/pk 186003030 186003184 186003155 186003331 186004438 186004866 4.6 x 50 mm Column 3.5 µm 1/pk 186003031 186003053 186003042 186003332 186004439 186004867 4.6 x 75 mm Column 3.5 µm 1/pk 186003032 186003185 186003043 186003333 — — 4.6 x 100 mm Column 3.5 µm 1/pk 186003033 186003054 186003044 186003334 186004440 186004868 4.6 x 150 mm Column 3.5 µm 1/pk 186003034 186003055 186003045 186003335 186004441 186004869 4.6 x 250 mm Column 3.5 µm 1/pk 186003943 186003963 186003964 186003965 — 186004870

2.1 x 10 mm Guard 5 µm 2/pk 1860030623 1860030803 1860030713 1860033663 1860044423 186006584 C and olumns 2.1 x 20 mm IS Column 5 µm 1/pk 186003107 186003186 186003156 186003336 — — 2.1 x 30 mm Column 5 µm 1/pk 186003129 186003187 186003157 186003337 186004443 186006587 2.1 x 50 mm Column 5 µm 1/pk 186003108 186003011 186002999 186003338 186004444 186006588 2.1 x 100 mm Column 5 µm 1/pk 186003109 186003012 186003002 186003339 186004445 186006589 2.1 x 150 mm Column 5 µm 1/pk 186003110 186003013 186003003 186003340 186004446 186006590 onsumables 3.0 x 20 mm Guard 5 µm 2/pk 1860030634 1860030814 1860030724 1860033674 — 186006585 3.0 x 20 mm IS Column 5 µm 1/pk 186003130 186003188 186003158 186003341 — — 3.0 x 30 mm Column 5 µm 1/pk 186003111 186003189 186003159 186003342 — — 3.0 x 50 mm Column 5 µm 1/pk 186003131 186003190 186003160 186003343 186004447 186006591 3.0 x 100 mm Column 5 µm 1/pk 186003132 186003191 186003004 186003344 186004448 186006592 3.0 x 150 mm Column 5 µm 1/pk 186003112 186003014 186003005 186003345 — — 3.0 x 250 mm Column 5 µm 1/pk 186003133 186003192 186003161 186003346 — — 4.6 x 20 mm Guard 5 µm 1/pk 1860030644 1860030824 1860030734 1860033684 1860044494 186006586 4.6 x 20 mm IS Column 5 µm 1/pk 186003134 186003193 186003162 186003347 — — 4.6 x 30 mm Column 5 µm 1/pk 186003135 186003194 186003163 186003348 186004450 — 4.6 x 50 mm Column 5 µm 1/pk 186003113 186003015 186003006 186003349 186004451 186006593 4.6 x 75 mm Column 5 µm 1/pk 186003114 186003195 186003007 186003350 — — 4.6 x 100 mm Column 5 µm 1/pk 186003115 186003016 186003008 186003351 186004452 186006594 4.6 x 150 mm Column 5 µm 1/pk 186003116 186003017 186003009 186003352 186004453 186006595

4.6 x 250 mm Column 5 µm 1/pk 186003117 186003018 186003010 186003353 186004454 186006596  See page 132 for product information. 3 Requires 2.1 x 10 mm Universal Sentry Guard Holder, Part No. WAT097958 4 Requires 3.0 x 20 mm/4.6 x 20 mm Universal Sentry Guard Holder, Part No. WAT046910

171 XBridge Columns Method Validation Kits Each Method Validation Kit contains 3 columns, each from a different batch.

Dimension Particle Size C18 C8 Shield RP18 Phenyl HILIC Amide 2.1 x 50 mm XP 2.5 µm 186006197 186006203 186006209 186006215 186006221 186006227 2.1 x 100 mm XP 2.5 µm 186006198 186006204 186006210 186006216 186006222 186006228 2.1 x 150 mm XP 2.5 µm 186006757 186006760 186006763 186006776 186006779 186006782 3.0 x 50 mm XP 2.5 µm 186006199 186006205 186006211 186006217 186006223 186006229 3.0 x 100 mm XP 2.5 µm 186006200 186006206 186006212 186006218 186006224 186006230 3.0 x 150 mm XP 2.5 µm 186006758 186006761 186006774 186006777 186006780 186006783 2.1 x 150 mm XP 2.5 µm 186006201 186006207 186006213 186006219 186006225 186006231 3.0 x 150 mm XP 2.5 µm 186006202 186006208 186006214 186006220 186006226 186006232 4.6 x 150 mm XP 2.5 µm 186006759 186006762 186006775 186006778 186006781 186006784 2.1 x 100 mm 3.5 µm 186003766 186003777 186003788 186003799 — — 3.0 x 100 mm 3.5 µm 186003767 186003778 186003789 186003800 — — 3.0 x 150 mm 3.5 µm 186003768 186003779 186003790 186003801 — — 4.6 x 100 mm 3.5 µm 186003769 186003780 186003791 186003802 — — 4.6 x 150 mm 3.5 µm 186003770 186003781 186003792 186003803 — — 2.1 x 150 mm 5 µm 186003771 186003782 186003793 186003804 — — 3.0 x 100 mm 5 µm 186003772 186003783 186003794 186003805 — — 3.0 x 150 mm 5 µm 186003773 186003784 186003795 186003806 — — 4.6 x 100 mm 5 µm 186003774 186003785 186003796 186003807 — — 4.6 x 150 mm 5 µm 186003775 186003786 186003797 186003808 — —

4.6 x 250 mm 5 µm 186003776 186003787 186003798 186003809 — —  onsumables See page 132 for product information.

XSelect Analytical Columns Particle CSH Fluoro- CSH Phenyl- Dimension Type Qty. CSH C HSS C HSS C SB HSS T3 HSS PFP HSS CN Size 18 Phenyl Hexyl 18 18 2.1 x 30 mm XP Column 2.5 µm 1/pk 186006100 186006112 186006124 186006136 186006160 186006148 186006172 186006184 olumns and C 2.1 x 30 mm XP Column 2.5 µm 3/pk 176002594 176002602 176002610 176002618 176002634 176002626 176002642 176002650 2.1 x 50 mm XP Column 2.5 µm 1/pk 186006101 186006113 186006125 186006137 186006161 186006149 186006173 186006185 2.1 x 50 mm XP Column 2.5 µm 3/pk 176002595 176002603 176002611 176002619 176002635 176002627 176002643 176002651 2.1 x 75 mm XP Column 2.5 µm 1/pk 186006102 186006114 186006126 186006138 186006162 186006150 186006174 186006186 2.1 x 75 mm XP Column 2.5 µm 3/pk 176002596 176002604 176002612 176002620 176002636 176002628 176002644 176002652 2.1 x 100 mm XP Column 2.5 µm 1/pk 186006103 186006115 186006127 186006139 186006163 186006151 186006175 186006187 2.1 x 100 mm XP Column 2.5 µm 3/pk 176002597 176002605 176002613 176002621 176002637 176002629 176002645 176002653 nalytical HPLC C 2.1 x 150 mm XP Column 2.5 µm 1/pk 186006727 186006730 186006733 186006736 186006742 186006739 186006745 186006748 A 2.1 x 150 mm XP Column 2.5 µm 3/pk 176002891 176002893 176002895 176002897 176002901 176002899 176002903 176002905 3.0 x 30 mm XP Column 2.5 µm 1/pk 186006104 186006116 186006128 186006140 186006164 186006152 186006176 186006188 3.0 x 30 mm XP Column 2.5 µm 3/pk 176002598 176002606 176002614 176002622 176002638 176002630 176002646 176002654 3.0 x 50 mm XP Column 2.5 µm 1/pk 186006105 186006117 186006129 186006141 186006165 186006153 186006177 186006189 3.0 x 50 mm XP Column 2.5 µm 3/pk 176002599 176002607 176002615 176002623 176002639 176002631 176002647 176002655 3.0 x 75 mm XP Column 2.5 µm 1/pk 186006106 186006118 186006130 186006142 186006166 186006154 186006178 186006190 3.0 x 75 mm XP Column 2.5 µm 3/pk 176002600 176002608 176002616 176002624 176002640 176002632 176002648 176002656 3.0 x 100 mm XP Column 2.5 µm 1/pk 186006107 186006119 186006131 186006143 186006167 186006155 186006179 186006191 3.0 x 100 mm XP Column 2.5 µm 3/pk 176002601 176002609 176002617 176002625 176002641 176002633 176002649 176002657 3.0 x 150 mm XP Column 2.5 µm 1/pk 186006728 186006731 186006734 186006737 186006743 186006740 186006746 186006749 3.0 x 150 mm XP Column 2.5 µm 3/pk 176002892 176002894 176002896 176002898 176002902 176002900 176002904 176002906 4.6 x 30 mm XP Column 2.5 µm 1/pk 186006108 186006120 186006132 186006144 186006168 186006156 186006180 186006192 4.6 x 50 mm XP Column 2.5 µm 1/pk 186006109 186006121 186006133 186006145 186006169 186006157 186006181 186006193 4.6 x 75 mm XP Column 2.5 µm 1/pk 186006110 186006122 186006134 186006146 186006170 186006158 186006182 186006194 4.6 x 100 mm XP Column 2.5 µm 1/pk 186006111 186006123 186006135 186006147 186006171 186006159 186006183 186006195

4.6 x 150 mm XP Column 2.5 µm 1/pk 186006729 186006732 186006735 186006738 186006744 186006741 186006747 186006750  See page 129 for product information. 3 Requires 2.1 x 10 mm Universal Sentry Guard Holder, Part No. WAT097958 4 Requires 3.0 x 20 mm/4.6 x 20 mm Universal Sentry Guard Holder, Part No. WAT046910

172 www.waters.com/order XSelect Analytical Columns Cont. Particle CSH Fluoro- CSH Phenyl- Dimension Type Qty. CSH C HSS C HSS C SB HSS T3 HSS PFP HSS CN Size 18 Phenyl Hexyl 18 18 1.0 x 50 mm Column 3.5 µm 1/pk 186005249 186005304 186005359 186006376 186006417 186006458 186005842 186005901 1.0 x 100 mm Column 3.5 µm 1/pk 186005250 186005305 186005360 186006377 186006418 186006459 186005843 186005903 1.0 x 150 mm Column 3.5 µm 1/pk 186005251 186005306 186005361 186006378 186006419 186006460 186005844 186005904 2.1 x 10 mm Guard 3.5 µm 2/pk 1860052523 1860053073 1860053623 1860063883 1860064293 1860064703 1860058643 1860059243 2.1 x 20 mm IS Column 3.5 µm 1/pk 186005253 186005308 186005363 186006379 186006420 186006461 186005845 186005905 2.1 x 30 mm Column 3.5 µm 1/pk 186005254 186005309 186005364 186006380 186006421 186006462 186005846 186005906 2.1 x 50 mm Column 3.5 µm 1/pk 186005255 186005310 186005365 186006381 186006422 186006463 186005847 186005907 2.1 x 75 mm Column 3.5 µm 1/pk 186005644 186005646 186005645 186006382 186006423 186006464 186005848 186005908 2.1 x 100 mm Column 3.5 µm 1/pk 186005256 186005311 186005366 186006383 186006424 186006465 186005849 186005909 2.1 x 150 mm Column 3.5 µm 1/pk 186005257 186005312 186005367 186006384 186006425 186006466 186005850 186005910 3.0 x 20 mm Guard 3.5 µm 2/pk 1860052584 1860053134 1860053684 1860047644 1860047454 1860047824 1860058654 1860059254 3.0 x 20 mm IS Column 3.5 µm 1/pk 186005259 186005314 186005369 186006385 186006426 186006467 186005851 186005911 3.0 x 30 mm Column 3.5 µm 1/pk 186005260 186005315 186005370 186004765 186004746 186004783 186005852 186005912 3.0 x 50 mm Column 3.5 µm 1/pk 186005261 186005316 186005371 186004766 186004747 186004784 186005853 186005913 3.0 x 75 mm Column 3.5 µm 1/pk 186005647 186005649 186005648 186005642 186005643 176005641 186005854 186005914 3.0 x 100 mm Column 3.5 µm 1/pk 186005262 186005317 186005372 186004762 186004743 186004780 186005855 186005915 A

3.0 x 150 mm Column 3.5 µm 1/pk 186005263 186005318 186005373 186004763 186004744 186004781 186005856 186005916 C HPLC nalytical 4.6 x 20 mm Guard 3.5 µm 2/pk 1860052644 1860053194 1860053744 1860047694 1860047504 1860047874 1860058664 1860059264 4.6 x 20 mm IS Column 3.5 µm 1/pk 186005265 186005320 186005375 186006386 186006427 186006468 186005857 186005917 4.6 x 30 mm Column 3.5 µm 1/pk 186005266 186005321 186005376 186004771 186004752 186004789 186005858 186005918 4.6 x 50 mm Column 3.5 µm 1/pk 186005267 186005322 186005377 186004772 186004753 186004790 186005859 186005919 4.6 x 75 mm Column 3.5 µm 1/pk 186005268 186005323 186005378 186006387 186006428 186006469 186005860 186005920 4.6 x 100 mm Column 3.5 µm 1/pk 186005269 186005324 186005379 186004767 186004748 186004785 186005861 186005921 4.6 x 150 mm Column 3.5 µm 1/pk 186005270 186005325 186005380 186004768 186004749 186004786 186005862 186005922 C and olumns 4.6 x 250 mm Column 3.5 µm 1/pk — — — 186004770 186004751 186004788 186005863 186005923 2.1 x 10 mm Guard 5 µm 2/pk 1860052713 1860053263 1860053813 1860064043 1860064453 1860064863 1860058873 1860059473 2.1 x 20 mm IS Column 5 µm 1/pk 186005272 186005327 186005382 186006389 186006430 186006471 186005867 186005927 2.1 x 30 mm Column 5 µm 1/pk 186005273 186005328 186005383 186006390 186006431 186006472 186005868 186005928 2.1 x 50 mm Column 5 µm 1/pk 186005274 186005329 186005384 186006391 186006432 186006473 186005869 186005929 2.1 x 100 mm Column 5 µm 1/pk 186005275 186005330 186005385 186006392 186006433 186006474 186005871 186005931 2.1 x 150 mm Column 5 µm 1/pk 186005276 186005331 186005386 186006393 186006434 186006475 186005872 186005932 onsumables 3.0 x 20 mm Guard 5 µm 2/pk 1860052774 1860053324 1860053874 1860064054 1860064464 1860064874 1860058884 1860059484 3.0 x 20 mm IS Column 5 µm 1/pk 186005278 186005333 186005388 186006394 186006435 186006476 186005873 186005933 3.0 x 30 mm Column 5 µm 1/pk 186005279 186005334 186005389 186006395 186006436 186006477 186005874 186005934 3.0 x 50 mm Column 5 µm 1/pk 186005280 186005335 186005390 186006396 186006437 186006478 186005875 186005935 3.0 x 100 mm Column 5 µm 1/pk 186005281 186005336 186005391 186006397 186006438 186006479 186005877 186005937 3.0 x 150 mm Column 5 µm 1/pk 186005282 186005337 186005392 186006398 186006439 186006480 186005878 186005938 3.0 x 250 mm Column 5 µm 1/pk 186005283 186005338 186005393 186006399 186006440 186006481 186005879 186005939 4.6 x 20 mm Guard 5 µm 2/pk 1860052854 1860053404 1860053954 1860047744 1860047554 1860047924 1860058894 1860059494 4.6 x 20 mm IS Column 5 µm 1/pk 186005284 186005339 186005394 186006400 186006441 186006482 186005880 186005940 4.6 x 30 mm Column 5 µm 1/pk 186005286 186005341 186005396 186006401 186006442 186006483 186005881 186005941 4.6 x 50 mm Column 5 µm 1/pk 186005287 186005342 186005397 186004852 186004757 186004794 186005882 186005942 4.6 x 75 mm Column 5 µm 1/pk 186005288 186005343 186005398 186006402 186006443 186006484 186005883 186005943 4.6 x 100 mm Column 5 µm 1/pk 186005289 186005344 186005399 186006403 186006444 186006485 186005884 186005944 4.6 x 150 mm Column 5 µm 1/pk 186005290 186005345 186005400 186004773 186004754 186004791 186005885 186005945

4.6 x 250 mm Column 5 µm 1/pk 186005291 186005346 186005401 186004775 186004756 186004793 186005886 186005946  See page 129 for product information. 3 Requires 2.1 x 10 mm Universal Sentry Guard Holder, Part No. WAT097958 4 Requires 3.0 x 20 mm/4.6 x 20 mm Universal Sentry Guard Holder, Part No. WAT046910

173 XSelect Columns Method Validation Kits Each Method Validation Kit contains 3 columns, each from a different batch.

Dimension Particle Size CSH C18 CSH Fluoro-Phenyl CSH Phenyl-Hexyl HSS C18 HSS C18 SB HSS T3 HSS PFP HSS CN 2.1 x 50 mm XP 2.5 µm 186006233 186006239 186006245 186006251 186006263 186006257 186006815 186006275 2.1 x 100 mm XP 2.5 µm 186006234 186006240 186006246 186006252 186006264 186006258 186006816 186006276 2.1 x 150 mm XP 2.5 µm 186006785 186006788 186006791 186006794 186006800 186006797 186006803 186006806 3.0 x 50 mm XP 2.5 µm 186006235 186006241 186006247 186006253 186006265 186006259 186006817 186006277 3.0 x 100 mm XP 2.5 µm 186006236 186006242 186006248 186006254 186006266 186006260 186006818 186006278 3.0 x 150 mm XP 2.5 µm 186006786 186006789 186006792 186006795 186006801 186006798 186006804 186006807 4.6 x 50 mm XP 2.5 µm 186006237 186006243 186006249 186006255 186006267 186006261 186006273 186006279 4.6 x 100 mm XP 2.5 µm 186006238 186006244 186006250 186006256 186006268 186006262 186006274 186006280 4.6 x 150 mm XP 2.5 µm 186006787 186006790 186006793 186006796 186006802 186006799 186006805 186006808 2.1 x 100 mm 3.5 µm 186005538 186005549 186005560 186006406 186006447 186006488 186005890 186005950 3.0 x 100 mm 3.5 µm 186005539 186005550 186005561 186006407 186006448 186006489 186005891 186005951 3.0 x 150 mm 3.5 µm 186005540 186005551 186005562 186006408 186006449 186006490 186005892 186005952 4.6 x 100 mm 3.5 µm 186005541 186005552 186005563 186006409 186006450 186006491 186005893 186005953 4.6 x 150 mm 3.5 µm 186005542 186005553 186005564 186006410 186006451 186006492 186005894 186005954 2.1 x 150 mm 5 µm 186005543 186005554 186005565 186006411 186006452 186006493 186005895 186005955 3.0 x 100 mm 5 µm 186005544 186005555 186005566 186006412 186006453 186006494 186005896 186005956 3.0 x 150 mm 5 µm 186005545 186005556 186005567 186006413 186006454 186006495 186005897 186005957 4.6 x 100 mm 5 µm 186005546 186005557 186005568 186006414 186006455 186006496 186005898 186005958 4.6 x 150 mm 5 µm 186005547 186005558 186005569 186006415 186006456 186006497 186005899 186005959

4.6 x 250 mm 5 µm 186005548 186005559 186005570 186006416 186006457 186006498 186005900 186005960  See page 129 for product information.

XTerra Analytical Columns

Dimension Type Particle Size Qty. MS C18 MS C8 RP18 RP8 Phenyl

onsumables 1.0 x 50 mm Column 2.5 µm 1/pk 186000979 — — — — 2.1 x 15 mm Direct Connect 2.5 µm 1/pk 186000900 — — — — 2.1 x 20 mm Cartridge 2.5 µm 1/pk 186000586 — — — — 2.1 x 20 mm IS Column 2.5 µm 1/pk 186001921 186001922 — — — 2.1 x 30 mm Column 2.5 µm 1/pk 186000592 186000593 — — — 2.1 x 50 mm Column 2.5 µm 1/pk 186000594 186000595 — — — 3.0 x 20 mm Cartridge 2.5 µm 1/pk 1860005889 1860005899 — — — 3.0 x 20 mm IS Column 2.5 µm 1/pk 186001972 186001973 — — — 3.0 x 30 mm Column 2.5 µm 1/pk 186000596 186000597 — — — olumns and C 3.0 x 50 mm Column 2.5 µm 1/pk 186000598 186000599 — — — 3.9 x 20 mm IS Column 2.5 µm 1/pk 186001899 186001897 — — — 4.6 x 20 mm Cartridge 2.5 µm 1/pk 1860005909 1860005919 — — — 4.6 x 20 mm IS Column 2.5 µm 1/pk 186001889 186001890 — — — 4.6 x 30 mm Column 2.5 µm 1/pk 186000600 186000601 — — — 4.6 x 50 mm Column 2.5 µm 1/pk 186000602 186000603 — — 186001138 75 µm x 50 mm Capillary 3.5 µm 1/pk 186002198 — — — — 75 µm x 100 mm Capillary 3.5 µm 1/pk 186002199 — — — — 75 µm x 150 mm Capillary 3.5 µm 1/pk 186002200 — — — — 100 µm x 50 mm Capillary 3.5 µm 1/pk 186002210 — — — — nalytical HPLC C 100 µm x 100 mm Capillary 3.5 µm 1/pk 186002211 — — — —

A 100 µm x 150 mm Capillary 3.5 µm 1/pk 186002212 — — — — 150 µm x 50 mm Capillary 3.5 µm 1/pk 186002469 — — — — 150 µm x 100 mm Capillary 3.5 µm 1/pk 186002470 — — — — 150 µm x 150 mm Capillary 3.5 µm 1/pk 186002471 — — — — 300 µm x 50 mm Capillary 3.5 µm 1/pk 186002599 — — — — 300 µm x 100 mm Capillary 3.5 µm 1/pk 186002600 — — — — 300 µm x 150 mm Capillary 3.5 µm 1/pk 186002601 — — — — 1.0 x 50 mm Column 3.5 µm 1/pk 186000386 186000387 186000388 186000389 — 1.0 x 100 mm Column 3.5 µm 1/pk 186000390 186000391 186000392 186000393 — 1.0 x 150 mm Column 3.5 µm 1/pk 186000394 186000395 186000396 186000397 — 2.1 x 10 mm Guard 3.5 µm 2/pk 1860006323 1860006333 1860006343 1860006353 1860011883 2.1 x 15 mm Direct Connect 3.5 µm 1/pk 186001908 — — — — 2.1 x 20 mm Guard 3.5 µm 2/pk 1860006365 1860006375 1860006385 1860006395 1860011895 2.1 x 20 mm IS Column 3.5 µm 1/pk 186001923 186001924 186001925 186001926 — 2.1 x 30 mm Column 3.5 µm 1/pk 186000398 186000399 — — — 2.1 x 50 mm Cartridge 3.5 µm 1/pk 1860004989 1860004999 1860005009 1860005019 — 2.1 x 50 mm Column 3.5 µm 1/pk 186000400 186000401 186000402 186000403 186001179 2.1 x 100 mm Cartridge 3.5 µm 1/pk 1860005029 1860005039 1860005049 1860005059 — 2.1 x 100 mm Column 3.5 µm 1/pk 186000404 186000405 186000406 186000407 186001180 2.1 x 150 mm Cartridge 3.5 µm 1/pk 1860005069 1860005079 1860005089 1860005099 —

2.1 x 150 mm Column 3.5 µm 1/pk 186000408 186000409 186000410 186000411 186001181  See page 140 for product information.

174 www.waters.com/order 1 Requires 10 x 10 mm Cartridge Holder, Part No. 289000779 6 Requires End Connector Kit, Part No. WAT037525 2 Requires 19 x 10 mm Cartridge Holder, Part No. 186000709 7 Requires 7.8 x 10 mm Cartridge Holder, Part No. 186000708 3 Requires 2.1 x 10 mm Universal Sentry Guard Holder, Part No. WAT097958 8 Requires Inline Guard Cartridge Holder, Part No. PSS830008 4 Requires 3.0 x 20 mm/4.6 x 20 mm Universal Sentry Guard Holder, Part No. WAT046910 9 Requires Cartridge Column End Connector Kit, Part No. 700000117 5 Requires Cartridge Column Holder, Part No. 186000262 XTerra Analytical Columns Cont.

Dimension Type Particle Size Qty. MS C18 MS C8 RP18 RP8 Phenyl 3.0 x 20 mm Guard 3.5 µm 2/pk 1860006404 1860006414 1860006424 1860006434 1860011904 3.0 x 20 mm IS Column 3.5 µm 1/pk 186001974 186001975 186001976 186001977 — 3.0 x 30 mm Column 3.5 µm 1/pk 186000412 186000413 — — — 3.0 x 50 mm Cartridge 3.5 µm 1/pk 1860005106 1860005116 1860005126 1860005136 — 3.0 x 50 mm Column 3.5 µm 1/pk 186000414 186000415 186000416 186000417 186001141 3.0 x 100 mm Cartridge 3.5 µm 1/pk 1860005146 1860005156 1860005166 1860005176 — 3.0 x 100 mm Column 3.5 µm 1/pk 186000418 186000419 186000420 186000421 186001142 3.0 x 150 mm Column 3.5 µm 1/pk 186000422 186000423 186000424 186000425 186001143 3.0 x 150 mm Cartridge 3.5 µm 1/pk 1860005186 1860005196 1860005206 1860005216 — 3.9 x 20 mm Guard 3.5 µm 2/pk 1860006444 1860006454 1860006464 1860006474 1860011914 3.9 x 20 mm IS Column 3.5 µm 1/pk 186001900 186001898 186001902 186001901 — 3.9 x 50 mm Cartridge 3.5 µm 1/pk 1860008176 1860008186 — — 186001204 3.9 x 100 mm Cartridge 3.5 µm 1/pk 1860005226 1860005236 1860005246 1860005256 — 3.9 x 100 mm Column 3.5 µm 1/pk 186000426 186000427 186000428 186000429 186001177 3.9 x 150 mm Column 3.5 µm 1/pk — — — — 186001178 4.6 x 10 mm Guard 3.5 µm 2/pk 1860019278 — — — — 4.6 x 20 mm IS Column 3.5 µm 1/pk 186001891 186001892 186001893 186001894 — 4.6 x 30 mm Column 3.5 µm 1/pk 186000430 186000431 186001910 186001912 — 6 6 6 6 4.6 x 50 mm Cartridge 3.5 µm 1/pk 186000526 186000527 186000528 186000529 — A

4.6 x 50 mm Column 3.5 µm 1/pk 186000432 186000433 186000434 186000435 — C HPLC nalytical 4.6 x 100 mm Cartridge 3.5 µm 1/pk 1860005306 1860005316 1860005326 1860005336 186001138 4.6 x 100 mm Column 3.5 µm 1/pk 186000436 186000437 186000438 186000439 186001139 4.6 x 150 mm Cartridge 3.5 µm 1/pk 1860005346 1860005356 1860005366 1860005376 — 4.6 x 150 mm Column 3.5 µm 1/pk 186000440 186000441 186000442 186000443 186001140 4.6 x 250 mm Column 3.5 µm 1/pk 186001470 186001471 186001472 186001473 186001474 300 µm x 50 mm Capillary 5 µm 1/pk 186002602 — — — — 300 µm x 100 mm Capillary 5 µm 1/pk 186002603 — — — — 300 µm x 150 mm Capillary 5 µm 1/pk 186002604 — — — — 2.1 x 10 mm Guard 5 µm 2/pk 1860006483 1860006493 1860006503 1860006513 1860011923

2.1 x 15 mm Direct Connect 5 µm 1/pk 186001907 — — — — C and olumns 2.1 x 20 mm Guard 5 µm 2/pk 1860006525 1860006535 1860006545 1860006555 1860011935 2.1 x 20 mm IS Column 5 µm 1/pk 186001979 186001980 186001982 186001983 — 2.1 x 50 mm Cartridge 5 µm 1/pk 1860005389 1860005399 1860005409 1860005419 — 2.1 x 50 mm Column 5 µm 1/pk 186000446 186000447 186000448 186000449 186001185 2.1 x 100 mm Cartridge 5 µm 1/pk 1860005429 1860005439 1860005449 1860005459 — 2.1 x 100 mm Column 5 µm 1/pk 186000450 186000451 186000452 186000453 186001186 2.1 x 150 mm Cartridge 5 µm 1/pk 1860005469 1860005479 1860005489 1860005499 — 2.1 x 150 mm Column 5 µm 1/pk 186000454 186000455 186000456 186000457 186001187

2.1 x 250 mm Cartridge 5 µm 1/pk 1860005509 1860005519 1860005529 1860005539 — onsumables 2.1 x 250 mm Column 5 µm 1/pk 186000458 186000459 186000460 186000461 — 3.0 x 20 mm Guard 5 µm 2/pk 1860006564 1860006574 1860006584 1860006594 1860011944 3.0 x 20 mm IS Column 5 µm 1/pk 186001984 186001985 186001986 186001987 — 3.0 x 50 mm Cartridge 5 µm 1/pk 1860005546 1860005556 1860005566 1860005576 — 3.0 x 50 mm Column 5 µm 1/pk 186000462 186000463 186000464 186000465 186001148 3.0 x 100 mm Cartridge 5 µm 1/pk 1860005586 1860005596 1860005606 1860005616 — 3.0 x 100 mm Column 5 µm 1/pk 186000466 186000467 186000468 186000469 186001149 3.0 x 150 mm Cartridge 5 µm 1/pk 1860005626 1860005636 1860005646 1860005656 — 3.0 x 150 mm Column 5 µm 1/pk 186000470 186000471 186000472 186000473 186001150 3.0 x 250 mm Cartridge 5 µm 1/pk 1860005666 1860005676 1860005686 1860005696 — 3.0 x 250 mm Column 5 µm 1/pk 186000474 186000475 186000476 186000477 186001151 3.9 x 20 mm Guard 5 µm 2/pk 1860006604 1860006614 1860006624 1860006634 1860011954 3.9 x 20 mm IS Column 5 µm 1/pk 186001988 186001989 186001990 186001991 — 3.9 x 50 mm Cartridge 5 µm 1/pk 1860008156 1860008166 — — 1860012036 3.9 x 100 mm Column 5 µm 1/pk — — — — 186001183 3.9 x 150 mm Cartridge 5 µm 1/pk 1860005706 1860005716 1860005726 1860005736 — 3.9 x 150 mm Column 5 µm 1/pk 186000478 186000479 186000480 186000481 186001184 4.6 x 10 mm Guard 5 µm 2/pk 1860019208 1860014348 — — — 4.6 x 30 mm Column 5 µm 1/pk 186000878 186000879 186001909 186001911 — 4.6 x 20 mm IS Column 5 µm 1/pk 186001992 186001993 186001994 186001995 — 4.6 x 50 mm Cartridge 5 µm 1/pk 1860005746 1860005756 1860005766 1860005776 — 4.6 x 50 mm Column 5 µm 1/pk 186000482 186000483 186000484 186000485 186001144 4.6 x 100 mm Column 5 µm 1/pk 186000486 186000487 186000488 186000489 186001145 4.6 x 150 mm Cartridge 5 µm 1/pk 1860005786 1860005796 1860005806 1860005816 — 4.6 x 150 mm Column 5 µm 1/pk 186000490 186000491 186000492 186000493 186001146 4.6 x 250 mm Cartridge 5 µm 1/pk 1860005826 1860005836 1860005846 1860005856 —

4.6 x 250 mm Column 5 µm 1/pk 186000494 186000495 186000496 186000497 186001147  See page 140 for product information. 1 Requires 10 x 10 mm Cartridge Holder, Part No. 289000779 3 Requires 2.1 x 10 mm Universal Sentry Guard Holder, Part No. WAT097958 4 Requires 3.0 x 20 mm/4.6 x 20 mm Universal Sentry Guard Holder, Part No. WAT046910 5 Requires Cartridge Column Holder, Part No. 186000262 6 Requires End Connector Kit, Part No. WAT037525 7 Requires 7.8 x 10 mm Cartridge Holder, Part No. 186000708 8 Requires Inline Guard Cartridge Holder, Part No. PSS830008 175 9 Requires Cartridge Column End Connector Kit, Part No. 700000117 XTerra Columns Method Validation Kits Each Method Validation Kit contains 3 columns, each from a different batch.

Dimension Particle Size Qty. MS C18 MS C8 RP18 RP8 Phenyl 2.1 x 100 mm 3.5 µm 1/pk Custom 186000831 Custom 186000834 Custom 4.6 x 100 mm 3.5 µm 1/pk Custom 186000832 Custom 186000835 Custom 4.6 x 150 mm 3.5 µm 1/pk 186000826 Custom 186000861 Custom 186002234 2.1 x 150 mm 5 µm 1/pk 186000827 Custom Custom Custom Custom 3.9 x 150 mm 5 µm 1/pk 186000828 Custom Custom 186000836 Custom 4.6 x 150 mm 5 µm 1/pk 186000829 Custom 186000862 Custom 186002235

4.6 x 250 mm 5 µm 1/pk 186000830 186000833 186000863 Custom 186002236  See page 140 for product information.

GC PoraPak Porous Polymer Packing Type Polarity Surface Area (m2/g) Density (g/cm3) Single Temp Prog. Particle Size Mesh Qty. Part No. 50–80 20 g WAT027053 P Nonpolar 100–200 0.26 250 ˚C 80–100 20 g WAT027054 100–120 20 g WAT027055 50–80 20 g WAT027083 PS Nonpolar 100–200 0.26 250 ˚C 80–100 20 g WAT027084 100–120 20 g WAT027085 50–80 26 g WAT027059 Slightly nonpolar

onsumables Q 500–600 0.34 250 ˚C 80–100 26 g WAT027060 to moderate 100–120 26 g WAT027061 50–80 26 g WAT027089 Slightly QS 500–600 0.34 250 ˚C 80–100 26 g WAT027090 nonpolar to moderate 100–120 26 g WAT027091 50–80 24 g WAT027065 Moderate polar olumns and C R monomer: 450–600 0.32 250 ˚C 80–100 24 g WAT027066 vinyl pyrolidone 100–120 24 g WAT027067 50–80 26 g WAT027071 Moderate polar S monomer: 300–450 0.35 250 ˚C 80–100 26 g WAT027072 vinyl pyridine 100–120 26 g WAT027073 50–80 29 g WAT027047 Polar polar monomer: N 250–350 0.41 190 ˚C 80–100 29 g WAT027048 vinyl pyrrolidone nalytical HPLC C 100–120 29 g WAT027049 A Highly polar polar 50–80 31 g WAT027077 monomer: T 225–350 0.39 190 ˚C 80–100 31 g WAT027078 ethyleneglycol

dimethacrylate 100–120 31 g WAT027079  See page 159 for product information.

Sentry Guard Holders Table Guard-Pak Holder (50 mm and 75 mm long Cartridge Columns must use the Universal Guard Holder) Description Part No. Description Part No. Guard-Pak Holder WAT088141

Integrated Guard Holder (for Waters Cartridge Columns) WAT046905 Guard-Pak Holder Connector WAT080046 Universal Guard Holder (for any HPLC Column) WAT046910  See page 148 for product information. Sentry 2.1 mm Guard Holder Kit WAT097958 (includes guard holder and rigid connector) O-Ring Kit (2/pk) for Sentry 2.1 mm Guard Holder WAT097954 Rigid Connector for Sentry 2.1 mm Guard Holder WAT022681

2.1 x 20 mm Guard Holder 186000262  See page 148 for product information.

176 www.waters.com/order Waters Cartridge and Guard Column Guide Cartridge Fittings and Accessories Guard Columns These endfittings and accessories can be used with 4.6 mm, 4.0 mm, or 3.0 mm ID cartridges. Universal Sentry Guard Holder Kits (use on any column)

2 3

4 Universal Sentry 1 Guard Holder

Dimension Part No. 1. 30 mm Stand Alone Guard/Column shown with endfittings (endfittings not included) 2. Guard holder endfitting for use with 10 mm Integral Guard, Part No. PSS614108 2.1 x 10 mm WAT097958 3. 10 mm Integral Guard Column 2.1 x 20 mm 186000262 4. Column Coupler, Part No. PSS614102 (for use with cartridge columns only) 3.0 x 20 mm WAT046910 Extended End-Fitting for use 3.9 x 20 mm WAT046910 Inline Guard Cartridge Holder with 10 mm Guard Cartridges 4.6 x 20 mm WAT046910

 See page 147 for product information. 10 mm Guard Cartridge (4.6 mm ID) 10 mm Guard Cartridge (4.6 mm ID) Waters Spherisorb Cartridge

Extended End-Fitting for use with 10 mm Guard, Part No. PSS 614108

Inline Guard Cartridge Holder, Part No. PSS 830008 High-Performance Carbohydrate Analysis Cartridge Column A Description Dimension Part No. C HPLC nalytical Description Qty. Part No. High-Performance Carbohydrate Cartridge Column 4.6 x 250 mm WAT044355 (requires endfittings) Removable Column End-Fitting 2/pk PSS614100 Sentry Integrated Guard Holder Frit Assembly (2 µm) 5/pk PSS614103 — WAT046905

(for Waters cartridge columns) Frit Assembly (0.5 µm) 5/pk PSS614104  Column Coupler 2/pk PSS614102 See page 154 for product information. Long Tail End-Fitting 2/pk PSS614101 Extended End-Fitting for use with 10 mm Integral Guard 1/pk PSS614108

Nylon Column Plugs for storage of Complete Column 1/pk WAT015674 C and olumns Nylon Column Caps for storage of Replacement Cartridge Column 10/pk PSS614113

Inline 10 mm Guard Cartridge Holder Kit for use with above items — PSS830008  Did you know... See page 147 for product information. Product care and use information is available on our website!

Go to: www.waters.com, under "Services and Support" onsumables look for "User Guides" or go to www.waters.com/chemcu Cartridge Columns

Cartridge with End-Fittings

Integrated Guard Holder Cartridge with Integrated Guard Column

Sentry Guard Column

Cartridge End-Fitting Applicable Column Dimension Part No. 2.1 x 50 mm, 2.1 x 100 mm, 2.1 x 150 mm, 2.1 x 250 mm 700000117 3.0 x 50 mm, 3.0 x 100 mm, 3.0 x 150 mm, 3.0 x 250 mm WAT037525 3.9 x 50 mm, 3.9 x 100 mm, 3.9 x 150 mm, 3.9 x 250 mm WAT037525

4.6 x 50 mm, 4.6 x 100 mm, 4.6 x 150 mm, 4.6 x 250 mm WAT037525  See page 147 for product information. Integrated Sentry Guard Holder to go with Cartridge End-Fittings (see above)

177 Application-Specific Columns

Column Dimension Particle Size Qty. Part No. Alcohols, Carbohydrates, Fermentation Analysis, and Organic Acids Carbohydrate Analysis Column — — 1/pk WAT01520913 Dextro-Pak Cartridge Column 8 x 100 mm — 1/pk WAT085650 Fast Fruit Juice Analysis 7.8 x 150 mm 4 µm 1/pk WAT010639 Fast Fruit Juice Guard-Pak Inserts — — 10/pk WAT0152077 High-Performance Carbohydrate 3.9 x 20 mm 4 µm 2/pk WAT04689516 Sentry Guard Column IC-Pak Ion Exclusion 7.8 x 300 mm — 1/pk WAT010290 SAM I Reagent — — 5/pk WAT010873 SC-1011 Column 8 x 300 mm — 1/pk WAT034238 SC-1011P Pre-Column 6 x 50 mm — 1/pk WAT034244 SH-1011 8 x 300 mm — 1/pk WAT034236 SH-1011P Pre-Column 6 x 50 mm — 1/pk WAT034243 Silica-Pak HP Cartridge 8 x 100 mm 4 µm 1/pk WAT010932 Silica-Pak Cartridge 8 x 100 mm 10 µm 1/pk WAT010935 SP-0810 Column 8 x 300 mm — 1/pk WAT036954 SP-0810P Pre-Column 6 x 50 mm — 1/pk WAT04245 Sugar-Pak 1 Column 6.5 x 300 mm — 1/pk WAT085188 Sugar-Pak 1 Guard-Pak Inserts 3.9 x 300 mm — 10/pk WAT084038 KC-811 8 x 300 mm 1/pk WAT034298 KC-811 Pre-Column 6 x 50 mm — 1/pk WAT035501

Carbamate Analysis Column for Pesticides Carbamate Analysis 3.9 x 150 mm — 1/pk WAT035577 onsumables

Free Fatty Acid HP Column Free Fatty Acid HP 3.9 x 150 mm 4 µm 1/pk WAT011690

IC-Pak Anion and Cation Analysis Columns IC-Pak Anion 4.6 x 50 mm — 1/pk WAT007355 IC-Pak Anion HR 4.6 x 75 mm — 1/pk WAT026765 olumns and C IC-Pak Anion HC 4.6 x 150 mm — 1/pk WAT026770 IC-Pak Anion Guard-Pak Kit — — 1/pk WAT007357 (Guard-Pak Holder and 5 inserts) IC-Pak Anion Concentrator Inserts — — 5/pk WAT00735813 IC-Pak Anion Guard-Pak Inserts — — 5/pk WAT01055113 IC-Pak C M/D Column 3.9 x 150 mm — 1/pk WAT036570 IC-Pak C M/D Guard-Pak Inserts — — 10/pk WAT04425013 IC-Pak Cation Column 4.6 x 50 mm — 1/pk WAT007354 13 nalytical HPLC C IC-Pak Cation Guard Column 4.6 x 50 mm — 1/pk WAT007356

A IC-Pak Cation Concentrator Inserts, — — WAT010565 5/pk

Ion-Exclusion Columns IC-Pak Ion Exclusion Column 7.8 x 150 mm — 1/pk WAT010295 IC-Pak Ion Exclusion Column 7.8 x 300 mm — 1/pk WAT010290 IC-Pak Ion Exclusion Guard-Pak — — 10/pk WAT02077013 Inserts

Waters PAH Columns PAH Column 4.6 x 50 mm 3 µm 1/pk 186001260 PAH Column 0.32 x 150 mm 5 µm 1/pk 186001259 PAH Column 2.1 x 150 mm 5 µm 1/pk 186001261 PAH Column 2.1 x 250 mm 5 µm 1/pk 186001262 PAH Column 3.0 x 250 mm 5 µm 1/pk 186001263 PAH Column 4.6 x 150 mm 5 µm 1/pk 186001264

PAH Column 4.6 x 250 mm 5 µm 1/pk 186001265  See page 154 for product information. 7 Requires 7.8 x 10 mm Cartridge Holder, Part No. 186000708 13 Requires Guard-Pak Holder, Part No. WAT088141 16 Requires Sentry Guard Holder, Part No. WAT046905

178 www.waters.com/order Standards Standards Environmental Analysis Description Part No. Standard Description Part No. Neutrals Suitability Standard 186006360 10 mg/mL of each pesticide: atrazin, atrazin-desethyl,

cyanazin, sebuthylazin, simazin, terbuthylazin, hexazinone,

Reversed-Phase Standard 186006363 20 Pesticide  chlortoluron, diuron, linuron, metobromuron, metoxuron, 186006348 Mix Standard See page 310 for product information. monolinuron, metolachlor, methamidophos, acephate, methomyl, dicrotophos, buprofenzin, fenpropimorph in acetonitrile 20 mL Total 0.5 mg/L sodium hypochlorite and Chlorine— 186006556 ammonium chloride in water, 20 mL

Cation Analysis 0.5 mg/L Standard Description Part No.  See page 317 for product information. 125 mL Ammonium NH4 Ammonium as NH4 in water, 100 mg/L 78102 125 mL Ammonium N Ammonium as N in water, 100 mg/L 78104 125 mL Lithium Lithium in water, 100 mg/L 78110 Paired Ion Chromatography (PIC) Reagents 125 mL Calcium Calcium in water, 1000 mg/L K10 Standard Description Part No. 125 mL Magnesium Magnesium in water, 1000 mg/L K11 5 vials of tetrabutylammonium hydrogen sulfate. PIC A Low UV Reagent WAT084189 125 mL Potassium Potassium in water, 1000 mg/L K12 Workable UV range in mobile phase is 200+ nm 125 mL Sodium Sodium in water, 1000 mg/L K13 PIC B5 Low UV 5 vials of pentane sulfonic acid. WAT084198

125 mL Nitrite NO2 Nitrite as NO2 in water, 1000 mg/L K15 Reagent Workable UV range in mobile phase is 200+ nm  PIC B6 Low UV 5 vials of hexane sulfonic acid. See page 319 for product information. WAT084199 Reagent Workable UV range in mobile phase is 200+ nm A

PIC B7 Low UV 5 vials of heptane sulfonic acid. C HPLC nalytical WAT084282 Reagent Workable UV range in mobile phase is 200+ nm Ion Calibration Standards PIC B8 Low UV 5 vials of octane sulfonic acid. WAT084283 Standard Description Part No. Reagent Workable UV range in mobile phase is 200+ nm 5 vials of tetrabutylammonium phosphate. 125 mL Ammonia NH3 Ammonia as NH3 in water, 1000 mg/L 044 PIC Reagent A WAT085101 Workable UV range in mobile phase is 240+ nm 125 mL Ammonia N Ammonia as N in water, 1000 mg/L 045 5 vials of pentane sulfonic acid. 125 mL Bromide Bromide in water, 1000 mg/L 046 PIC Reagent B5 WAT085110 Workable UV range in mobile phase is 240+ nm 125 mL Chloride Chloride in water, 1000 mg/L 047 5 vials of hexane sulfonic acid. PIC Reagent B6 WAT085140 125 mL Fluoride Fluoride in water, 1000 mg/L 050 Workable UV range in mobile phase is 240+ nm C and olumns 125 mL Nitrate NO3 Nitrate as NO3 in water, 1000 mg/L 051-ERA 5 vials of heptane sulfonic acid. PIC Reagent B7 WAT085103 125 mL Nitrate N Nitrate as N in water, 1000 mg/L 052-ERA Workable UV range in mobile phase is 240+ nm 5 vials of octane sulfonic acid. 125 mL Nitrite N Nitrite as N in water, 1000 mg/L 053 PIC Reagent B8 WAT085142 Workable UV range in mobile phase is 240+ nm 125 mL Phosphate PO4 Phosphate as PO4 in water, 1000 mg/L 060 5 vials of dibutylammonium phosphate. 125 mL Phosphate P Phosphate as P in water, 1000 mg/L 061-ERA PIC Reagent D4 WAT085466 Workable UV range in mobile phase is 215+ nm 125 mL Sulphate Sulphate in water, 1000 mg/L 062 Kit WAT085144 125 mL Phosphorus Phosphorous in water, 1000 mg/L 063

PIC A WAT085101 onsumables 125 mL Silica Silica in water, 1000 mg/L 064 PIC Reagent Sample B5 WAT085110 125 mL Bromate Bromate in water, 1000 mg/L 065 Kit (One Each) B6 WAT085140 125 mL Chlorate Chlorate in water, 1000 mg/L 066 B7 WAT085103 125 mL Chlorite Chlorite in water, 1000 mg/L 067 B8 WAT085142

125 mL Perchlorate Perchlorate in water, 1000 mg/L 068

®  For use with Pico•Tag PITC Reagent WAT088120 See page 318 for product information. amino acid analysis method

SAM I Reagent 5/box; reagent for carbohydrate analysis WAT010873  See page 324 for product information. QC Trace Metal Standards Standard Description Part No. Standard Description Part No. 125 mL Aluminum Aluminum in nitric acid , 1000 mg/L 011 125 mL Manganese Manganese in nitric acid, 1000 mg/L 026 125 mL Antimony Antimony in nitric acid, 1000 mg/L 012 125 mL Mercury Mercury in nitric acid, 1000 mg/L 027 125 mL Arsenic Arsenic in nitric acid, 1000 mg/L 013 125 mL Molybdenum Molybdenum in nitric acid, 1000 mg/L 028 125 mL Barium Barium in nitric acid, 1000 mg/L 014 125 mL Nickel Nickel in nitric acid, 1000 mg/L 029 125 mL Beryllium Beryllium in nitric acid, 1000 mg/L 015 125 mL Potassium Potassium in nitric acid, 1000 mg/L 030 125 mL Boron Boron in nitric acid, 1000 mg/L 016 125 mL Selenium Selenium in nitric acid, 1000 mg/L 031 125 mL Cadmium Cadmium in nitric acid, 1000 mg/L 017 125 mL Silicon Silicon in nitric acid, 1000 mg/L 032 125 mL Calcium Calcium in nitric acid, 1000 mg/L 018 125 mL Silver Silver in nitric acid, 1000 mg/L 033 125 mL Chromium VI Chromium VI in water, 1000 mg/L 019 125 mL Sodium Sodium in nitric acid, 1000 mg/L 034 125 mL Chromium Chromium in nitric acid, 1000 mg/L 020 125 mL Strontium Strontium in nitric acid, 1000 mg/L 035 125 mL Cobalt Cobalt in nitric acid, 1000 mg/L 021 125 mL Thallium T halium in nitric acid, 1000 mg/L 036 125 mL Copper Copper in nitric acid, 1000 mg/L 022 125 mL Tin Tin in hydrochloric acid, 1000 mg/L 037 125 mL Iron Iron in nitric acid, 1000 mg/L 023 125 mL Titanium Titanium in hydrochloric acid, 1000 mg/L 038 125 mL Lead Lead in nitric acid, 1000 mg/L 024 125 mL Vanadium Vanadium in nitric acid, 1000 mg/L 039

125 mL Magnesium Magnesium in nitric acid, 1000 mg/L 025 125 mL Zinc Zinc in nitric acid, 1000 mg/L 040  See page 318 for product information. 179