Perform Rugged, Fast LC with Confidence. Agilent InfinityLab Poroshell 120 Columns for HPLC and UHPLC

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1 Perform Rugged, Fast LC with Confidence Agilent InfinityLab Poroshell Columns for HPLC and UHPLC

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3 A tradition of innovation and efficiency More than years ago, Agilent introduced the industry's first superficially porous column technology with Poroshell 3 for large molecules, followed by Poroshell for small molecules. Today, the InfinityLab Poroshell family has grown to include 8 chemistries including new phases for chiral and HILIC separations. Whether you use traditional HPLC systems or newer UHPLC systems, you will experience exceptional separation efficiency, and significantly boost performance and throughput from every LC in your lab. InfinityLab Poroshell columns A wide range of innovative chemistries supports optimal separation of a variety of analytes. Excellent lot-to-lot reproducibility: A proprietary, single-step porous shell process dramatically reduces tiny differences between lots and columns. A scalable family of particles:.9 μm,.7 μm, and 4 μm superficially porous particles enable you to get the best from your methods and all of your instruments. Easy method transfer between HPLC and UHPLC: Scalable particles and similar chemistries across ZORBAX and Poroshell products simplify transferability of methods. Fast method development: Up to 8 chemistries maximize selectivity so you can find the optimal separation for your sample. Long column life: Robust Poroshell particles and chemistries are stable at required pressures. UHPLC guard columns further extend the life of your analytical column. Superior peak shape: High-purity silica and advanced bonding chemistries reduce peak tailing and give you faster, more accurate results. Easy traceability An optional preprogrammed ID tag allows you to track column properties and usage parameters on your Agilent InfinityLab Series LC 3

4 Table of Contents What makes InfinityLab Poroshell columns different? Page How our superficially porous particles are made contributes to the excellent results you can achieve Which column should I choose? Page 7 Find the best column for your application Develop methods quickly and easily Page 8 8 chemistries provide a wide range of selectivities New columns for polar separations Page 3 Achieve the best possible retention with new HILIC columns New columns for chiral compounds Page Shorten run times for chiral separations Make every LC work harder Page 8 Perform high-speed, high-resolution separations on your current instruments Increase the flexibility of all your methods Page Now you can perform very fast, high-efficiency separations under the widest range of separation conditions USP method transfer, made simpler Page Easily move your legacy methods to InfinityLab Poroshell columns Ordering information Page 7 Part numbers and specifications To learn more about Agilent InfinityLab Poroshell columns, visit 4

5 What makes InfinityLab Poroshell columns different? InfinityLab Poroshell columns are based on superficially porous particle technology, which features a solid silica core and a porous outer layer. Compared to traditional totally porous particles of the same (or similar) size, Poroshell particles deliver higher chromatographic efficiencies and enable fast, highresolution separations. Poroshell superficially porous particles deliver high efficiency and reproducibility through several advantages over totally porous particles: Uniform particles with smooth surfaces ensure smaller particle size distributions Short analyte diffusion path in/out of the porous layer Narrow particle size distributions allow more uniform packing of column particles How is a Poroshell particle made? Agilent uses a unique manufacturing process for Poroshell particles. Specifically, we imize the number of manufacturing steps to ensure maximum particle and chromatographic reproducibility. Step : Make the solid core InfinityLab Poroshell cores have a very smooth surface and a uniform particle size both of which contribute to a tight overall particle size distribution, improving analytical performance and reproducibility. Step : Apply the porous shell At Agilent, we apply the porous shell in one single step. This unique process delivers more column-to-column reproducibility than other vendors columns. Step 3: Apply the bonded phase A wide range of robust bonded phases enable analysis of a variety of analytes in different LC modes, to solve even your toughest separation challenges.

6 Reproducible performance lot to lot and year to year imizes workflow disruption The simpler the manufacturing process, the more consistent the column A single-step shell process creates a highly reproducible column, as you can see in this comparison of five lots of InfinityLab Poroshell columns.. years B B8 B4 B B4 InfinityLab Poroshell EC-C8, 3. x mm,.7 µm (p/n 997-9) from five different lots A scalable family of particles for faster method transfer InfinityLab Poroshell columns are available in three different particle dimensions allowing you to choose the size that best fits your separation needs, and is most compatible with your LC systems. Because the different particles are engineered with a consistent core-to-particle size ratio, you can easily transfer a method developed on one particle size to any of the others. Quality you can count on All Agilent columns are designed and manufactured end-to-end by Agilent, undergoing multiple QC tests to tight specifications. Take a closer look at our stringent manufacturing processes at

7 Which InfinityLab Poroshell column should I choose? InfinityLab Poroshell columns are available in three particle sizes to fit all your separation needs.9 µm Highest UHPLC performance.7 µm UHPLC performance at lower pressures 4 µm Improved HPLC performance System Max pressure (bar) Typical LC instrument We recommend UHPLC (very low dispersion) -+ Agilent 9 Infinity II InfinityLab Poroshell.9 µm InfinityLab Poroshell.7 µm UHPLC (moderate-low dispersion) - Agilent Infinity II Agilent Infinity II Prime InfinityLab Poroshell.7 µm InfinityLab Poroshell 4 µm HPLC 4 - Agilent Infinity II InfinityLab Poroshell 4 µm InfinityLab Poroshell.7 µm With 8 different chemistries, InfinityLab Poroshell columns provide a range of selectivities that makes your method development fast and easy. Best all around Best for low-ph mobile phases Best for high-ph mobile phases Best for polar compounds (HILIC) Best for alternative selectivity Best for Chiral separations EC-C8.9,.7, 4 µm SB-C8.7 µm HPH-C8.9,.7, 4 µm HILIC.9,.7, 4 µm Bonus-RP.7 µm Chiral-T.7 µm EC-C8.9,.7, 4 µm SB-C8.7 µm HPH-C8.7, 4 µm HILIC-Z.7um PFP.9,.7, 4 µm Chiral-V.7 µm HILIC-OH.7 µm Phenyl-Hexyl.9,.7, 4 µm Chiral-CD.7 µm SB-Aq.7 µm Chiral-CF.7 µm EC-CN.7 µm 7

8 Develop methods quickly and easily with the right selectivities Best all around: InfinityLab Poroshell EC-C8 and EC-C8 Agilent InfinityLab Poroshell EC- C8 provides a similar chemistry to the ZORBAX Eclipse Plus phase for ideal separation of a wide variety of analytes. Agilent InfinityLab Poroshell EC-C8 is less retentive for faster analysis of nonpolar compounds. Mobile phase: % CH 3 CN, 4% H O Flow rate:.8 ml/ Temperature: ºC Detection: 4 nm Sample: µl of RRLC checkout sample (p/n 88-9), alkylphenones InfinityLab Poroshell EC-C8, 3. x mm,.7 µm InfinityLab Poroshell EC-C8, 3. x mm,.7 µm EC-C8 is an excellent starting point. Use EC-C8 for less retention with a variety of samples. Best for low-ph mobile phases: InfinityLab Poroshell SB-C8 and SB-C8 SB-C8 and SB-C8 are made using bulky silanes that sterically protect the siloxane bond. Acid labile endcapping reagents are not used. The result is vastly improved column life and extraordinary chemical and temperature stability at ph -. A lack of endcapping also provides a different selectivity to the endcapped EC-C8 and EC-C8. InfinityLab Poroshell SB-C8. x mm x.7 µm Column: InfinityLab Poroshell SB-C8,. x mm,.7 µm Mobile phase: A: Acid in H O B: CH 3 CN Flow rate:.79 ml/ Gradient: Time %B Temperature: 4 ºC Source: 3 ºC, L/, psi, -3 V Acquisition: SIM Neg (9, 3, 93**, 89, 47, 44) **caffeine is not detected in Neg mode Sample: 3 µl of 3 µg/ml each of GA, GC, EGC, C, Caf, EC, EGCG, GCG, ECG, CG in H O/ CH 3 CN.% acetic acid.% formic acid S/N= S/N= Samples: Gallic acid Gallocatechin Epigallocatechin Catechin Caffeine Epicatechin Epigallocatechin gallate Gallocatechin gallate Epicatechin gallate Catechin gallate Agilent InfinityLab Poroshell SB chemistry provides exceptional peak shape and column stability at low-ph. A separation of catechins in green tea demonstrates the value of screening acidic modifiers to enhance LC/MS detection sensitivity. Learn more at 8

9 Best for high-ph mobile phases: InfinityLab Poroshell HPH-C8 and HPH-C8 HPH-C8 and HPH-C8 incorporate hybrid InfinityLab Poroshell particle technology to provide high-ph stability. Hybrid particle technology improves overall particle ruggedness at extended ph, enabling long lifetimes and fewer column changes. The selectivity of the HPH chemistries is very similar to EC-C8 and EC-C8, making method transfer simple. Instrument: Infinity II Binary LC Mobile phase: A: mm Ammonium bicarbonate adjusted to ph. in water B: Acetonitrile Flow rate:.4 ml/ Gradient: Time % B 9. Sample:. Methyl salicylate. 4 Chlorocinnamic acid 3. Acetophenone 4. Quinine. Nortryptyline. Heptanophenone 7. Amitriptyline InfinityLab Poroshell HPH-C8,. x mm,.7 µm Injection Injection, Injection, After, injections at ph, InfinityLab Poroshell HPH-C8 showed no change in performance. A robust method development process is critical to ensuring that your method is long lasting, stable, and reliable. Because the retention and selectivity of ionizable compounds can change significantly with varying ph, it is becog standard practice to employ low, medium, and high-ph analyses during method development. Flow rate: ml/ UV detection: 4 nm Gradient: Time %Buffer %MeCN Sample:. Procainamide. Caffeine 3. Acetyl salicylic acid 4. Hexanophenone deg.. Dipyrimadole. Diltiazem 7. Diflunisal 8. Hexanophenone InfinityLab Poroshell HPH-C8, 4. x mm,.7 µm 7 ph 3, mm NH 4 CH 3 CO ph 4.8, mm NH 4 CH 3 CO ph, mm NH 4 HCO Here, a method using low, medium, and high ph separates the same mixture of acids, bases, and neutrals. The highest resolution for all compounds was obtained under higher-ph conditions; therefore, high ph would be the best choice going forward. 9

10 Best for alternative selectivity: InfinityLab Poroshell Bonus-RP, PFP, and Phenyl-Hexyl Alternate selectivities allow you to explore various conditions to find optimal separation of your analytes of interest. The Phenyl-Hexyl phase provides alternate selectivity to C8 phases, particularly for analytes containing aromatic groups. Bonus-RP provides unique mid-ph selectivity with an embedded amide linkage in the alkyl chain. InfinityLab Poroshell Bonus-RP,. x mm,.7 µm InfinityLab Poroshell Phenyl-Hexyl,. x mm,.7 µm InfinityLab Poroshell EC-C8,. x mm,.7 µm Instrument: Infinity II Binary LC Mobile phase: A: mm NH 4 HCO, ph 3.8 B: MeOH Flow rate:.4 ml/ Temperature: 4 C Detection: nm Gradient: % B to 3% B/ Sample:. Atenolol. Acebutolol. Pindolol. Propranolol 3. Nadolol 7. Alprenolol 4. Metoprolol Analysis of beta blockers: a comparison of InfinityLab Poroshell phases. This challenging separation demonstrates different selectivities across bonded phases. Overall, the Bonus-RP phase delivered the best peak shape and resolution.

11 PFP is a pentafluorophenyl ligand, which provides an orthogonal separation mechanism with C8 chemistries. PFP phases can separate analytes based on small differences in structure, substitution, and steric access to polar moieties. The resulting selectivity for positional isomers, halogenated compounds, and polar analytes is particularly useful when analyzing complex mixtures. InfinityLab Poroshell PFP 4. x mm,.7 µm InfinityLab Poroshell 4EC-C8 4. x mm, 8.7 µm 4 3 7,8 3 3, , , InfinityLab Poroshell Phenyl-Hexyl 4. x mm,.7 µm , InfinityLab Poroshell EC-C8 4. x mm,.7 µm Mobile phase: A: Water (.% acetic acid) B: Acetonitrile Flow rate: ml/ Positional isomers ( compounds) Sample:. 3,4 Dimethoxyphenol., Dimethoxyphenol 3. 3, Dimethoxyphenol 4., Difluorophenol.,4 Difluorophenol.,3 Difluorophenol 7. 3,4 Difluorophenol 8. Degradation product, dimethoxyphenol 9. 3, Dimethylphenol., Dimethylphenol., Dichlorophenol. 4 Chloro 3 methyl phenol 3. 4 Chloro methyl phenol 4. 3,4 Dichlorophenol. 3, Dichlorophenol

12 Alternate selectivity for polar compounds: InfinityLab Poroshell SB-Aq SB-Aq is a proprietary alkyl reversed-phase chemistry designed to retain hydrophilic and other compounds when using highly aqueous mobile phases including % water. InfinityLab Poroshell SB-Aq, 3 mm,.7 µm Standards Cranberry cocktail Grapefruit juice Orange juice Eluent: mm Potassium phosphate buffer, ph. Injection volume: μl Flow rate:. ml/ Temperature: C Detector: DAD, at nm Sample:. Tartaric acid. Quinic acid 3. Malic acid 4. Citric acid. Fumaric acid Agilent InfinityLab Poroshell EC-CN offers very different selectivity compared to C8 for mixtures of polar, mid-polar and non-polar compounds. The EC-CN phase can also be used in normal phase mode for retention of polar compounds. InfinityLab Poroshell EC-CN,. mm,.7 μm 7 3, 4,, 3, InfinityLab Poroshell EC-C8,. mm,.7 μm , 8,, Alternate selectivity is observed when comparing InfinityLab Poroshell EC-CN and EC-C8 phases for the analysis of explosives using EPA 833A method parameters. Instrument: Sample concentration: Eluent: Injection volume: μl Flow rate: Isocratic: Agilent 9 Infinity LC μg/ml of each compound A, H O; B, CH 3 OH.4 ml/ % B Temperature: C Detector: 4 nm Sample: Explosive residues. HMX. RDX 3.,3,-Trinitrobenzene 4.,3-Dinitrobenzene. Nitrobenzene.,4,-Trinitrotoluene 7. -Ao-4,-dinitrotoluene 8.,4-Dinitrotoluene 9. Tetryl. 4-Ao-,-dinitrotoluene.,-Dinitrotoluene. -Nitrotoluene 3. 4-Nitrotoluene 4. 3-Nitrotoluene Learn more at

13 Analyze challenging polar compounds with confidence Hydrophilic interaction chromatography (HILIC) is gaining popularity due to its ability to retain and separate small polar analytes using a standard LC system and common reversed-phase solvents. Advantages of this technique include: Simplicity: Just change the column and adjust the method to analyze highly polar compounds Better retention of polar analytes than reversed-phase methods MS-compatible eluents and easy setup Improved ionization and sensitivity in MS mode InfinityLab Poroshell HILIC,. x mm,.7 µm x Mobile phase: Acetonitrile: mm NH 4 HC, ph 3. (9:) Flow rate:.7 ml/ Temperature: C Pressure: 4 bar System: 9 Infinity II LC and 4 Triple Quadrupole LC/MS Sample:. 4 Aobenzoic acid. Nicotinamide 3. Riboflavin 4. Nicotinic acid Counts (%) vs. Acquisition Time () InfinityLab Poroshell HILIC eliates the need for ion-pair reagents, such as the hexane sulfonic acid that is typically used in mobile phases for separating B vitas. It also improves LC/MS compatibility and retention. 3

14 Retain highly charged compounds, including inorganic salts: t New InfinityLab Poroshell HILIC-Z This Agilent column innovation features a novel zwitterionic stationary phase bonded to Poroshell particles. Advantages include: High peak capacity and wide polarity range A PEEK-lined column option for excellent peak shape and recovery of challenging compounds Robust performance High ph stability: Up to ph at 3 ºC; Temperature stability to 8 ºC at ph 7 Tolerates samples with high salt or buffer content Compatible with MS-friendly buffers (< mm salt content); low MS bleed InfinityLab Poroshell HILIC-Z. x mm,.7 µm x Phenylalanine Leucine Isoleucine Methionine Tyrosine Valine Proline Alanine Cysteine Threonine Glycine Serine Glutamic Acid Aspartic Acid Histidine Arginine Lysine Cystine Mobile phase: A: mm ammonium formate in H, ph3 B: 9: ACN/H with mm ammonium formate, ph3 Gradient: % B to 7% B over, return to % B Flow rate:.8 ml/ Temp 3 ºC MS detection: Agilent MS-QQQ, MS SIM mode Excellent retention, peak shape, and sensitivity for underivatized ao acids by LC/MS. 4

15 Separate polar compounds with alternate selectivity: t New InfinityLab Poroshell HILIC-OH New poly-hydroxy fructan phase bonded to Poroshell particles Fast, high efficiency separations Excellent retention of polar compounds Offers alternate selectivity to HILIC and HILIC-Z phases InfinityLab Poroshell HILIC-OH. x mm,.7 μm B B B3 B B B9 C B InfinityLab Poroshell HILIC-Z. x mm,.7 μm B,B3 B B B B9 C B InfinityLab Poroshell HILIC. x mm,.7 μm B B B C B3 B B B9 InfinityLab HILIC-Z and HILIC-OH columns offer a wide range of alternate selectivity for polar analyte separations. This separation of water soluble vitas demonstrates selectivity differences across HILIC phases using the same method conditions. Separations can be optimized on each column by altering the method conditions. Mobile phase: A: mm Ammonium Acetate +.% Acetic Acid (ph ~4.) in H O, B: CH 3 CN. ml/, 87% B for, 87-% B in 4, 3 re-equilibration Flow rate: μl injection, 4 ºC Temperature: 4 ºC Detector: UV detection at nm, 8 Hz Samples: thiae (B) riboflavin (B) niacin/nicotinic acid (B3) pyridoxine/pyridoxol (B) folate/folic acid (B9 4-aobenzoic acid/paba (B) cyanocobala (B) asorbic acid (C)

16 Perform fast, high-resolution chiral separations Now you don t have to compromise on your chiral separations t New InfinityLab Poroshell Chiral columns Agilent chiral columns combine superficially porous particles with innovative chiral stationary phases to deliver: Higher performance and speed, compared to totally porous chiral stationary phases Ruggedness and reliability with proven Agilent Poroshell particle technology Increased sample throughput and lab productivity with more efficient chiral separations Perform chiral separations in less than utes using your existing LC systems Superficially porous particles provide higher efficiency and sharper peak shapes. Throughput is dramatically improved with shorter runtimes, compared to fully porous particle columns. Traditional Chiral Separation totally porous particle Chirobiotic V (4. x mm, µm) 3 4 Eluent: /./., MeOH/HOAc/NH 4 OH Flow Rate:. ml/ Detector: UV at nm Sample:. D-(+)-Amphetae. L-(-)-Amphetae 3. D-(+)-Methamphetae 4. L-(-)-Methampheta Agilent Poroshell Chiral Separation superficially porous particle InfinityLab Poroshell Chiral-V (4. x mm,.7 µm) 3 4 Separation of amphetaes in under utes Eluent: /./., MeOH/HOAc/NH 4 OH Flow Rate:. ml/ Detector: UV at nm Sample:. D-(+)-Amphetae. L-(-)-Amphetae 3. D-(+)-Methamphetae 4. L-(-)-Methampheta

17 Separate nearly any chiral compound: Four chemistries across four different LC modes offer a wide range of selectivities Column Chemistry Chiral Selector (bonded chemistry) Typical LC Mode Typical Applications InfinityLab Poroshell Chiral-CF Derivatized cyclofructan (CF) Polar Organic (PO) Normal Phase (NP) Primary aes Primary aes InfinityLab Poroshell Chiral-CD Hydroxypropylated-β-cyclodextrin Reversed Phase (RP) Polar Organic (PO) Stimulants, fungicides, t-boc ao acids Complex molecules Polar Ionic (PI) Basic pharmaceuticals (various) InfinityLab Poroshell Chiral-V Vancomycin (macrolide antibiotic) Reversed Phase (RP) Aes, profens Polar Organic (PO) Complex neutral molecules Polar Ionic (PI) Beta blockers, hydroxyl acids InfinityLab Poroshell Chiral-T Teicoplanin (macrolide antibiotic) Reversed Phase (RP) Ao acids, hydroxyl acids, profens Polar Organic (PO) Hydantoins, benzodiazepines InfinityLab Poroshell Chiral CF InfinityLab Poroshell Chiral CD F * NH F * N N N OH -(-napthyl)ethylae flutriafol Rs= 3.3 Rs= Column: 4. x mm,.7 µm Mobile phase: /4/.3/.: Acetonitrile/Methanol/Acetic Acid/TEA Flow Rate:. ml/ Detection: UV 8 nm Column: 4. x mm,.7 µm Mobile phase: 3/7: Acetonitrile/ mm Ammonium Formate (ph 3.) Flow Rate:. ml/ Detection: UV 3 nm InfinityLab Poroshell Chiral V InfinityLab Poroshell Chiral T N H OH * O propanolol Cl NH O OH p-chlorophenylalanine * Rs=.4 Rs= Column: 4. x mm,.7 µm Mobile phase: /./.: Methanol/Acetic Acid/ Ammonium Hydroxide Flow Rate:. ml/ Detection: UV 3 nm 3 4 Column: 4. x mm,.7 µm Mobile phase: 3/7: Acetonitrile/ mm Ammonium Formate (ph 3.) Flow Rate:. ml/ Detection: UV nm 7

18 Make every LC work harder Long column lifetimes provide added value You can count on InfinityLab Poroshell particles to be robust under the most demanding operating conditions. In addition, InfinityLab Poroshell.7 µm and 4 µm columns are manufactured with µm inlet frits to resist plugging, even with the dirtiest samples. InfinityLab Poroshell EC-C8 3. x mm,.7 µm Pressure End press 8 Plates Injections Robust InfintyLab Poroshell.9 µm columns last up to five times longer than other SPP columns. Longer column lifetimes reduce operating costs by preventing expensive disruption and re-work due to column failure. Efficiency (N) Instrument: Injection volume: µl Solvent A: Solvent B: Flow Rate: Infinity RRLC (SL) Water with. % TFA MeCN with.8 % TFA ml/ Time %B Sample: Precipitated plasma: parts plasma, 7 parts :8 water: MeCN with.% formic acid with part diflunisal in : water: MeCN µg/ml (final concentration diflunisal µg/ml) shaken and allowed to settle utes..% formic acid aids in the precipitation process. Not centrifuged and not filtered Disruption/re-work needed per column failure Analyst cost of labor per hour Number of columns required to run year s worth of samples Column Lifetime, Other SPP Columns [4 hours] $ InfinityLab Poroshell.9 µm Columns : hours utes, Other SPP Columns Agilent InfinityLab Poroshell.9 µm Columns Learn more at 8

19 Get more from every LC system in your lab System Max pressure (bar) Typical LC instrument We recommend UHPLC (very low dispersion) -+ Agilent 9 Infinity II InfinityLab Poroshell.9 µm InfinityLab Poroshell.7 µm UHPLC (moderate-low dispersion) - Agilent Infinity II Agilent Infinity II Prime InfinityLab Poroshell.7 µm InfinityLab Poroshell 4 µm HPLC 4 - Agilent Infinity II InfinityLab Poroshell 4 µm InfinityLab Poroshell.7 µm Ultrafast separations: InfinityLab Poroshell.9 µm InfinityLab Poroshell.9 µm columns generate lower pressures than many other sub- µm superficially porous columns. That means you can use high flow rates to drive ultrafast separations without exceeding the pressure rating of your UHPLC instruments. InfinityLab Poroshell EC-C8,. x mm,.9 µm 3 Pressure bar Instrument: Agilent 9 Infinity II Mobile phase: A:.% formic acid in water 3 Run time.3 Mobile phas B: Acetonitrile Gradient: 8-% B in.3 Flow rate:. ml/ Temperature: ºC Detection: 8 Hz Sample: μl of. mg/ml each of protocatechuic acid, DOPAC, PABA, vanillic acid, syringic acid, salicylic acid An ultrafast UHPLC separation with baseline resolution of aromatic acids using an InfinityLab Poroshell.9 µm column together with a high flow rate. 9

20 UHPLC performance at HPLC pressures: InfinityLab Poroshell.7 µm Achieve up to 9% or more of the efficiency you would expect from a sub- μm totally porous particle column, but at HPLC pressures. This enables you to maximize the separation performance and analytical throughput of your HPLC systems. You can also use shorter columns at higher flow rates to achieve fast separations dramatically enhancing productivity and decreasing cost per sample. InfinityLab Poroshell EC-C8, 3. x mm,.7 µm N =,3, Pressure = 8 bar ZORBAX Eclipse Plus C8, 3. x mm,.8 µm Mobile phase: % Acetonitrile, 4% water Flow rate:.8 ml/ Injection volume: 4 µl Temperature: ºC Detection: DAD Sig = 4,4 nm Ref = 3, nm Sample: RRLC checkout sample (p/n 88-9) spiked with µl mg/ml thiourea in water: acetonitrile (:3) N = 7,9, Pressure = 38 bar Use a.7 µm Poroshell particle column to obtain >9% of the efficiency at half the pressure of a.8 µm totally porous particle column. Improved HPLC performance: InfinityLab Poroshell 4 µm InfinityLab Poroshell 4 μm columns offer nearly double the efficiency of totally porous μm particles. That means you can improve separations performance by simply replacing the µm totally porous column of your legacy methods, without changing column dimensions or requiring significant method development. What's more, alignment between ZORBAX and Poroshell chemistries makes this an easy transition for immediate improvement in your analysis. InfinityLab Poroshell EC-C8, 4. x mm, 4 µm 8 4 ZORBAX Eclipse Plus C8, 4. x mm, µm 8 4 Initial Pressure 3 bar Initial Pressure 9 bar Rs=.3 Rs=. 3 4 Improved results with no change to the method conditions, other than the column used. In this example, direct replacement with an InfinityLab Poroshell 4 μm column decreased peak width by % and improved resolution compared to the μm totally porous column. 4 4 W / =. W / =. Mobile phase: A: Water B: Acetonitrile Gradient for 4. x mm columns: Time () %A Temperature: 3 C Flow rate:. ml/ Injection volume: µl Detection: UV, 3 nm Sample:. Notoginsenoside R. Ginsenoside Rg 3. Ginsenoside Re 4. Ginsenoside Rb

21 Agilent InfinityLab Agilent InfinityLab is designed to provide you with the highest efficiency in your LC and LC/MSD workflows regardless of application area. InfinityLab is an optimized portfolio of LC instruments, mass selective detectors, columns, and supplies designed to work together in perfect harmony. Combined with Agilent OpenLAB software and Agilent CrossLab Services, Agilent provides you with end-to-end solutions and support to make every day more productive. More at: InfinityLab LC Series From routine analysis to cutting-edge research, the Agilent InfinityLab LC Series provides the broadest portfolio of LC systems and solutions for any application and budget to achieve highest efficiency. Affordable efficiency The Agilent Infinity II LC is an affordable, high-quality integrated LC system, putting you on the fast track to efficiency. Everyday efficiency in every way The Agilent Infinity II LC is the trusted platform with the broadest instrument choice, taking you to the next level of efficiency. Agilent Infinity II LC Agilent Infinity II LC Agilent 9 Infinity II LC The benchmark in efficiency The Agilent 9 Infinity II LC embodies the next generation of LC, giving you the ultra-high performance to achieve maximum efficiency. InfinityLab Supplies Save time and imize troubleshooting InfinityLab Quick Connect fittings Now you can be sure you re getting a perfect column connection, from every user, every time with simple-to-use, finger-tight connections. Quick- Connect fittings are reusable for more than reconnections and stable up to 3 bar. InfinityLab Quick Turn fittings For instrument connections that are in confined spaces, you can rely on spring-loaded, Agilent InfinityLab Quick Turn fittings for a perfect connection, every time.

22 . Flexible method handling from UHPLC to legacy to LC/MS Flexibility to choose longer columns for higher resolution As the column gets longer, resolution improves and pressure increases. Maximize resolution by running longer columns on an Agilent InfinityLab Series or 9 UHPLC system with InfinityLab Poroshell columns rated to, or 3 bar. InfinityLab Poroshell EC-C8, 4. x mm,.7 µm - P = 3 bar Mobile phase: % Methanol, 7% water Flow rate: ml/ Temperature: 44 ºC InfinityLab Poroshell EC-C8, 4. x mm - - P = 38 bar InfinityLab Poroshell EC-C8, 4. x mm 4 3 P = 8 bar Balancing column length, resolution, and analysis time are important for any separation to obtain the optimal separation in the fastest runtime, thereby maximizing lab throughput and productivity.

23 Simple transfer of legacy methods Easily transfer legacy methods to InfinityLab Poroshell to gain the separation and throughput benefits of superficially porous particles. Alignment between ZORBAX and Poroshell chemistries simplifies this transition. In the example below, comparable chemistries are used and all method conditions were kept the same, except the gradient time which was adjusted for the shorter column. ZORBAX Eclipse Plus C8, 4. x mm, µm 8 4 bar InfinityLab Poroshell EC-C8, 4. x mm,.7 µm 3 bar Flow rate: ml/ Time %B Flow rate: ml/ Time %B More than 4x increase in throughput Mobile phase: A:.% Formic acid in water B:.% Formic acid in acetonitrile Sample:. Sulfadiazine. Sulfathiazole 3. Sulfapyridine 4. Sulfamerazine. Sulfamethazine. Sulfamethazole 7. Sulfamethoxypyridazine 8. Sulfachloropyridazine 9. Sulfamethoxazole. Sulfadimethoxine In this example, a legacy method is transitioned from a ZORBAX Eclipse Plus C8 column to a comparable Poroshell EC-C8 phase. Comparable resolution is acheived in a much shorter runtime on the InfinityLab Poroshell column, improving analytical throughput by 4x. Increase flow rate for fast UHPLC separations Take advantage of the pressure capabilities of InfinityLab Poroshell columns to speed up your separations..9 µm and.7 µm InfinityLab Poroshell columns have lower backpressure than competitive columns, allowing you to increase the flow rates to maximize throughput. Upper pressure limits of bar, bar and 3 bar for InfinityLab Poroshell columns allow you to get the most out of your UHPLC instruments. InfinityLab Poroshell EC-C8 3. x mm,.7 µm Flow rate =. ml/, P = 3 bar 4 Flow rate =.7 ml/, P = 433 bar Mobile phase: A: %,.% Formic acid B: 3% Methanol isocratic Flow rate: see chromatograms Injection volume: µl Temperature: ºC Detection: Sig =, 4 nm, Ref = Off Sample:. Saccharin. Caffeine 3. p-hydroxybenzoic acid 4. Aspartame. Dehydroacetic acid. Benzoic acid Flow rate =. ml/, P = 9 bar 4 3

24 Get the best performance from your LC/MS methods High-efficiency InfinityLab Poroshell columns are ideally suited to improve the resolution and sensitivity of your LC/MS methods. Use a. or 3 mm internal diameter (ID) column for the best flow rate compatibility with your MS.. A 4. mm Ammonium formate InfinityLab Poroshell 4.% Formic HPH-C8, acid, 3. ph x 3., MeOH, mm, 3 4 µm C 3. ml/. A. A 3. 8 Time % B 4. mm Ammonium formate 7 4. mm Ammonium formate 9 4.% Formic acid, ph 3., MeOH, 3 C 4.%. Formic acid, ph 3., MeOH, 3 C. 3. ml/ 3. ml/ Time %. 8 Time % B B Sample: B Strychnine mm Ammonium Bicarbonate 9. Alprazolam MDMA ph., MeOH, 3 C 3 ml/ Sample: 4. Amphetae B Sample: 4. B. 7. Strychnine. Trazodone 7 8. Strychnine 4 mm Ammonium Bicarbonate 4 8. Alprazolam. Meperidine mm. Ammonium Bicarbonate. Alprazolam 3. ph., MeOH, 3 C MDMA 7. Verapamil ph., MeOH, 3 C 9 3. MDMA 3 ml/ 4. Amphetae 8. Methadone. ml/. 4.. Amphetae Trazodone 9. Proadifen 4. Trazodone. Meperidine.Diazepam Meperidine Verapamil. THC Verapamil 8. Methadone Methadone Proadifen 9..Diazepam Proadifen.Diazepam. THC THC Counts Counts Counts Counts Counts Counts Excellent resolution for a separation of drugs of abuse by LC/MS using the InfinityLab Poroshell HPH column. The high-ph stability of HPH allows the method to be run at ph. where an increase in retention and MS response for basic analytes is observed. LC Column Navigator tool Find a more efficient replacement for your current column or get recommendations for a new column, based on method parameters. 4

25 USP method transfer, made simpler Modernize a USP method for naproxen tablets: 4.x faster analysis on Agilent InfinityLab Poroshell columns at HPLC pressures This naproxen separation demonstrates how easy it can be to convert a method to InfinityLab Poroshell columns without changing the flow rate or mobile phase. The first chromatogram shows a USP analysis on an Agilent ZORBAX Eclipse Plus C8 column. In the second and third chromatograms, the InfinityLab Poroshell EC-C8 4 µm columns ( mm and mm) provide greater efficiency and speed than the original method as easy, drop-in replacements with reasonable pressures for HPLC. In the fourth chromatogram, the InfinityLab Poroshell EC-C8.7 µm column ( mm) provides greater efficiency and resolution at nearly times the speed of the original method. A shorter InfinityLab Poroshell EC-C8 column ( mm) in the fifth chromatogram still meets the requirements for efficiency and resolution, but is 4. times faster than the µm column. ZORBAX Eclipse Plus C8, 4. x mm, µm USP Prescribed Column mau 4 3 mau 4 3 N =, InfinityLab Poroshell, 4. x mm (L) 4 µm mau 4 3 N = 9,4 InfinityLab Poroshell, 4. x mm (L), 4 µm N = 3,8 L/dp = 3, 8 9 x Faster, same backpressure 98 bar L/dp =, InfinityLab Poroshell EC-C8, 4. x mm (L),.7 µm mau 3 N =,4 x Faster N =, bar bar x Efficiency L/dp = 37, µl injection Rs = 7. L/dp = 37,37 P = 38 bar InfinityLab Poroshell EC-C8, 4. x mm (L),.7 µm System suitability method requirement: N > 4,, Rs >. Mobile phase: :49: MeCN:H O:acetic acid Flow rate:. ml/ Sample:. Naproxen. Butyrophenone mau 3 N =,8 N =, 4.x Faster Alignment of InfinityLab Poroshell and ZORBAX chemistries means that selectivities are very similar between column families making it easy to transfer your legacy methods..7 µl injection Rs =. L/dp =8,8 P= 33 bar

26 Transfer of USP methods USP methods are widely used in pharmaceutical drug products and raw materials testing. These methods can be updated by making adjustments that follow the recommendations in USP chapter <> (updated August 4). Modifications outside these ranges are considered method changes, and typically require method revalidation. Parameters for System Suitability USP37-NF3S Isocratic Particle size (dp) L/dp: -% to +% or Column length (L) N: -% to +% Column inner diameter Flow rate Injection volume Column temperature Mobile phase ph Salt concentration Ratio of components in mobile phase Wavelength of UV-Visible detector Flexible, w/ constant linear velocity Based on dp Additional adjustments: ±%, provided N decreases % May be adjusted as far as is consistent with precision and detection limits ± o C ±. units Within ±% if the permitted ph variation is met Minor component ( %): ±3% relative, but cannot exceed ±% absolute May only adjust or component in ternary mixtures No changes allowed L (mm) dp (µm) L/dp % N % compliant 3, %,39 % Yes 4 37, % 9,4 79% Yes 4, 83% 3,8 4% Yes.7 37,37 3%,4 98% Yes.7 8,9 %,8 % Yes L L3 L7 L L L43 L L9 EC-C8 HILIC EC-C8 EC-CN Phenyl-Hexyl PFP Bonus-RP SB-Aq SB-C8 HPH-C8 SB-C8 HPH-C8

27 Ordering information InfinityLab Poroshell column specifications InfinityLab Poroshell Family Pore Size Temp. Limits ph Range Endcapped Carbon Load Surface Area Best all around Best for low-ph mobile phases Best for high-ph mobile phases EC-C8 Å C.-8. Double % 3 m/g EC-C8 Å C.-8. Double % 3 m/g SB-C8 Å 9 C.-8. No 9% 3 m/g SB-C8 Å 8 C.-8. No.% 3 m/g HPH-C8 Å C 3.-. Double Proprietary 9 m/g HPH-C8 Å C 3.-. Double Proprietary 9 m/g HILIC Å C.-8. N/A N/A 3 m/g Best for polar compounds (HILIC) HILIC-Z Å 8 C 3.-. Proprietary Proprietary 3 m/g HILIC-OH Å 4 C.-7. Double Proprietary 3 m/g Bonus-RP Å C.-9. Triple 9.% 3 m/g PFP Å C.-8. Double.% 3 m/g Best for alternative selectivity Phenyl-Hexyl Å C.-8. Double 9% 3 m/g SB-Aq Å 8 C.-8. No Proprietary 3 m/g EC-CN Å C.-8. Double 3.% 3 m/g Chiral-T Å 4 C.-7. Proprietary Proprietary 3 m/g Best for Chiral separations Chiral-V Å 4 C.-7. Proprietary Proprietary 3 m/g Chiral-CD Å 4 C Proprietary Proprietary 3 m/g Specifications represent typical values only Chiral-CF Å 4 C Proprietary Proprietary 3 m/g Particle Pressure Limit.9 µm 3 bar.7 µm bar* 4 µm bar * unless otherwise noted Easy-to-use InfinityLab UHPLC Fast Guards High-performance Agilent InfinityLab UHPLC Fast Guards are designed for use with fast LC columns. They connect directly to the column inlet; no extra hardware is needed. Agilent UHPLC Fast Guards are available in most InfinityLab Poroshell chemistries giving you confidence that the guard column will not adversely affect your separations. 7

28 InfinityLab Poroshell.9 µm columns: Highest UHPLC performance Size (mm) EC-C8 EC-C8 Phenyl-Hexyl HPH-C8 PFP HILIC 3. x x x x x x Note: All InfinityLab Poroshell.9 µm columns are supplied with a pre-programmed Column ID. InfinityLab Poroshell.9 µm UHPLC Fast Guards: Extend column lifetime even further Size (mm) EC-C8 EC-C8 Phenyl-Hexyl HPH-C8 PFP HILIC 3. x x Note: Guards supplied as 3/pk. InfinityLab Poroshell.7 µm Chiral columns: High-resolution separations of chiral compounds Size (mm) Chiral-CF Chiral-CD Chiral-V Chiral-T. x x x x x x Note: Rated to 4 bar. InfinityLab Poroshell Chiral columns are not available with pre-programmed column ID tags. InfinityLab Poroshell.7 µm HILIC columns: Best for polar separations Size (mm) HILIC HILIC-OH HILIC-Z HILIC-Z (PEEK-lined). x x x x x x x x x Note: InfintyLab Poroshell HILIC-OH has a 4 bar maximum pressure limit and is not available with a pre-programmed Column ID. 8

29 InfinityLab Poroshell.7 µm columns: UHPLC performance at lower pressures Size (mm) EC-C8 EC-C8 SB-C8 SB-C8 HPH-C8 HPH-C8 4. x x x x x x x 937-3* 3. x x 97-3* 3. x x x x x 937-9*. x x 97-9*. x x x Size (mm) Phenyl-Hexyl Bonus-RP PFP SB-Aq EC-CN 4. x x x x x x x x x Note: InfinityLab Poroshell.7 µm columns have a bar/9, psi pressure limit unless otherwise noted. InfinityLab Poroshell.7 μm columns are not supplied with a pre-programmed Column ID as standard. To order with Column ID, suffix the part number with T (example T). *These columns have a maximum pressure limit of, bar InfinityLab Poroshell.7 µm Fast Guards: Extend column lifetime even further Size (mm) EC-C8 EC-C8 Phenyl-Hexyl SB-C8 SB-C8 HPH-C8 HPH-C8 4. x x x Size (mm) Bonus-RP PFP SB-Aq EC-CN HILIC HILIC-Z 4. x x x Note: Guards supplied as 3/pk. 9

30 InfinityLab Poroshell 4 µm columns: Improved HPLC performance Size (mm) EC-C8 EC-C8 Phenyl-Hexyl HPH-C8 HPH-C8 PFP HILIC 4. x x x x x x x x x x x x Note: InfinityLab Poroshell 4 µm columns have a bar/9, psi pressure limit. InfinityLab Poroshell 4 μm columns are not supplied with a pre-programmed Column ID as standard. To order with Column ID, suffix the part number with T (example 997-9T) InfinityLab Poroshell 4 µm Fast Guards: Extend column lifetime even further Size (mm) EC-C8 HPH-C8 HPH-C8 4. x x x Note: Guards supplied as 3/pk. Agilent AdvanceBio LC Columns: Part of the InfinityLab Family Find more information at The flexibility to achieve full biomolecule characterization requires a tool box of columns appropriate for large molecule separations. Whether you are analyzing intact proteins, protein fragments, peptide digests, ao acids, or oligonucleotides, Agilent offers a comprehensive range of columns in different pore sizes and particle morphologies designed for these complex separations. All are supported by expert technical support and application chemists around the globe. Agilent AdvanceBio LC columns enhance the accuracy and speed of your biomolecule characterization. 3

31 Method Validation Kits Contain 3 HPLC columns with the same column type (bonded phase, particle size, configuration), but from different manufacturing lots. Size (mm) Particle Size (µm) EC-C8 EC-C8 Phenyl-Hexyl SB-C8 SB-C8 SB-Aq Bonus-RP 4. x K K K K K K K 4. x K 997-9K 997-9K 897-9K 897-9K K 998-9K 4. x K K K K K K K 3. x K K K K K K K 3. x K 997-3K 997-3K 897-3K 897-3K K 998-3K 3. x K K K K K K K. x K K K K K K K. x K 977-9K 977-9K 877-9K 877-9K K 978-9K. x K K K K K K K Size (mm) Particle Size (µm) EC-C8 EC-C8 Phenyl-Hexyl PFP HILIC 4. x K 997-9K 997-9K K 997-9K 4. x K K K K K 4. x K 997-9K 997-9K K 997-9K 4. x K K K K K 3. x K 997-3K 997-3K K 997-3K 3. x K K K K K 3. x K 997-3K 997-3K K 997-3K 3. x K K K K K. x 4 7-9K 7-9K 7-9K 7-48K 7-9K. x K K K K K. x K 977-9K 977-9K K 977-9K. x K K K K K Don t see a validation kit for your column part number? Validation kits can be assembled for most columns in the Agilent LC column portfolio. Simply contact your Agilent sales specialist or send your request to the Agilent customer center in your country, indicating the part number for which you need a validation kit. An Agilent representative will send you a quote within - business days. 3

32 Agilent Sample Preparation portfolio Sample preparation is an essential component of successful chromatography for improved analytical robustness and to prevent operational downtime due to contaation of columns and instrumentation. Agilent offers a complete line of sample prep products for filtration, SPE, SPME, SLE and sample processing devices. Learn more at: Maximize uptime with end-to-end support You can trust Agilent CrossLab service experts to deliver valuable insights and keep your instruments running at top performance. Our industry-leading services tailored to meet your needs include instrument transition, application consulting, repairs, preventive maintenance, compliance verification, and education. Ask us how we can support your laboratory today. Learn more at: Learn more: Buy online: U.S. and Canada agilent_inquiries@agilent.com Europe info_agilent@agilent.com Asia Pacific inquiry_lsca@agilent.com This information is subject to change without notice. Agilent Technologies, Inc. 7, 8 Published in the USA, March, EN