Agilent Poroshell 120 Columns for HPLC and UHPLC PERFORM RUGGED, FAST LC WITH CONFIDENCE

Transcription

1 Agilent Poroshell Columns for HPLC and UHPLC PERFRM RUGGED, FAST LC WITH CNFIDENCE

2 AGILENT PRSHELL CLUMNS CAN MAKE EVERY LC AND LC/MS IN YUR LAB WRK EVEN HARDER We choose Poroshell because of its rugged performance Poroshell provides reliably excellent performance it s the new standard in our lab For complicated samples, which I face most, Poroshell columns save me a lot of time Poroshell is my go-to column QUTES ARE FRM PRSHELL USERS Poroshell columns provide exceptional efficiency for standard HPLC, and significantly boost the performance you ll get from all instruments, whether you have older bar or newer bar UHPLC instruments. These next-generation columns take the technology introduced with our Poroshell columns to the next level giving you higher throughput and resolution for a wider range of small molecules and peptides than ever before. Their advanced features include: Extraordinary lot-to-lot reproducibility Poroshell columns are manufactured using a proprietary single-step porous shell process that dramatically reduces ute differences between columns and lots Comparable speed and resolution to sub- µm columns and significant improvements over μm columns with much lower backpressure take HPLC and UHPLC performance to a new level of flexibility and efficiency Superior peak shape especially at ph -7 for faster, more accurate results Long column life Poroshell columns use a standard μm frit, and resist plugging with dirty samples Up to TWELVE Chemistries, depending on the particle size, including SB-C and SB-C for low ph applications and Poroshell HPH-C and HPH-C for high ph applications Easy method transfer to ZRBAX bonded phases and within the Poroshell family, for highest productivity from lab to lab, around the world UHPLC guard column options that reduce your operating expenses by extending the life of Poroshell columns Scalability within the Poroshell Family with column options in μm and.7 μm configurations for optimal performance for your method

3 Here is what you ll find in this brochure What makes Poroshell columns different? Unique processes for making our superficially porous particles and bonded phases contribute to the excellent results you can achieve Page A family of particle sizes and bonded phases for flexible selectivity and scalability Poroshell offers both μm and.7 μm columns scalable to and from ZRBAX phases Page Making your HPLC work harder Yes you can achieve Fast LC performance at HPLC pressures Page Making every LC and LC/MS work harder Perform high-speed, high-resolution separations on your current instruments Page 9 Increasing the flexibility of your UHPLC methods Now you can perform very fast, high-efficiency separations under the widest range of separation conditions Page Easy method transfer Save time and money by moving methods from μm or. μm columns to Poroshell columns Page Solutions to frustrating throughput and resolution problems How Poroshell columns help you meet the challenges you face every day Page New options for proteins and peptides Achieve faster peptide mapping and protein separations with Poroshell columns Page Infinitely better liquid chromatography Introducing Agilent Infinity II Series LC systems Page Specifications and ordering information Page To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell

4 HIGH EFFICIENCY AT LWER PRESSURES PRVEN CLUMN-T-CLUMN CNSISTENCY THAT S THE PRSHELL DIFFERENCE A key feature of Agilent Poroshell columns is their superficially porous microparticulate column packing. Poroshell columns offer both μm and.7 μm particle sizes with scalable performance, consisting of a solid silica core with a porous shell. This unique configuration gives you all the performance advantages of smaller totally porous particles, but with lower backpressures.. μm totally porous Agilent Poroshell.7 μm Agilent Poroshell μm. μm.7 μm.7 μm. μm Pore size Å, ideal for small molecules. μm.7 μm μm How a Poroshell particle is made In order to create the best column for small molecule separations, we completely reinvented our superficially porous particle technology. Specifically, we imized the manufacturing steps involved to ensure maximum particle and chromatographic reproducibility. STEP Make the solid core Poroshell column cores have a very smooth surface and a uniform particle size which contributes to a tight overall particle size distribution. As a result, you get a more tightly packed column bed and higher efficiency than with totally porous particles. STEP Apply the porous shell At Agilent, we apply the porous shell in one single step similar to the coacervation technique used to make traditional ZRBAX columns. This unique single-step process delivers higher yields and more column-tocolumn reproducibility than competitors. STEP Apply the bonded phase The family of Agilent Poroshell phases is expanding to align with the ZRBAX family for method development flexibility and assured scalability. See the following pages for details on all of the Poroshell phases that are available to you.

5 A comparison of particle size distributions between totally porous and Poroshell particles This graph demonstrates that Agilent Poroshell columns have the tightest final particle size distribution a direct result of starting with a tight core size distribution. Number Agilent Poroshell.7 µm Agilent. µm RX SIL Agilent. µm RX SIL Agilent. µm RX SIL The tight final particle size distribution is a result of starting with a tight core size distribution. This helps imize diffusion and results in high efficiency of superficially porous columns (µm) The standard measure of particle size distribution is the 9/ ratio, which should be below. The ZRBAX totally porous particles (. μm,. μm, and. μm) all have an acceptable particle size distribution. However, the Poroshell particle has a % tighter particle size distribution, which substantially improves column efficiency. Agilent Poroshell (.7 µm) LN B Agilent ZRBAX. µm Agilent ZRBAX. µm Agilent ZRBAX. µm %. µm.7 µm.7 µm.9 µm 9%. µm. µm. µm. µm 9%/% ratio..7.. Reproducible performance from lot to lot, year after year Poroshell particles are made with a proprietary porous particle manufacturing process, invented by Agilent. Instead of traditional multilayering, Poroshell columns are manufactured using a single-step coacervation process that produces a more consistent final particle and more reliable chromatographic results. 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 lot-to-lot comparison of five lots. Agilent Poroshell EC-C,. x mm,.7 µm P/N from Different Lots B B B B B To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell

6 A VARIETY F BNDED PHASES MEANS YU NEVER HAVE T CMPRMISE N SELECTIVITY Poroshell columns are made at the same facility as Agilent s industryleading ZRBAX column family. The bonding chemistries used with Poroshell columns mirror those of all ZRBAX columns, giving you the advantages of easier method transfer and assured scalability from lab to lab, around the world. All the selectivities you need to perfect your separation AN EXCELLENT FIRST CHICE H H N N N N N N N N Poroshell EC-C and EC-C (USP L)* You can count on this high-performance phase to deliver excellent peak shape and resolution for acids, bases, and neutrals. This chemistry is very similar to the ZRBAX Eclipse Plus phase, for easy method transferability. Exceptional peak shape, efficiency, resolution and lifetime. Tip: Select the C phase first, and use the C phase for less retention with a variety of samples. H N N N N N N A variety of published methods from Agilent use Poroshell, allowing for easy method development. H H H H H H N H C N C N N Methods easily convert from [ZRBAX] Eclipse Plus columns to Poroshell we use them for all methods. QUTES FRM ACTUAL TXICLGY LAB USERS

7 Poroshell Phenyl-Hexyl (USP L)* This phase offers an alternative selectivity for phenyl groups, and is very similar to ZRBAX Eclipse Plus Phenyl-Hexyl for easy method transfer. H C H H N CH CH NEW Poroshell PFP* F Provides an alternative selectivity for halogenated compounds and polar analytes. H CH H HIGH ph APPLICATINS NEW Poroshell HPH-C and HPH-C The silica in this special chemistry has been modified with a proprietary process to increase stability at high ph levels. H H H H Poroshell Bonus-RP (USP L) Bonus-RP is polar-embedded to improve peak shape for basic compounds at low- and mid-ph. This phase is the same as ZRBAX Bonus-RP. S H H H H F F H H N CH CH H C N CH H H H N CH H LW ph APPLICATINS StableBond SB-C and SB-C (USP L) StableBond performs well with acids, bases, and neutrals with superior lifetime at low ph. What s more, these phases transfer readily from ZRBAX SB-C and ZRBAX SB-C phase chemistries. H Poroshell HILIC* With its unbonded silica, Poroshell HILIC enables you to retain and separate small polar analytes. Poroshell EC-CN (USP L) Similar to ZRBAX Eclipse XDB-CN, this cyano phase simplifies method transfer. H H N N H H H H N H C H CH HCl Poroshell SB-Aq This proprietary phase provides an alternate selectivity option, and is ideal for polar compounds and high aqueous conditions. Its chemistry is the same as ZRBAX SB-Aq. H H H H H H H H H H H H H * Now available in μm and.7 μm particle sizes. Visit agilent.com/chem/discoverporoshell for videos, Application Notes, and more or to order now 7 7

8 Agilent Poroshell EC-C and Poroshell SB-C provide different selectivity for optimizing separations Agilent Poroshell SB-C. x mm,.7 µm P/N 97- Mobile phase: % H, % CH CN Flow rate: ml/ Temperature: ºC MS acquisition: Dynamic MRM Compound Precursor ion Fragmentor voltage Anandamide (AEA) Palmitoylethanolamide (PEA) -Arachidonoylglycerol (-AG) 79 leoylethanolamide (EA) MS Source: Gas temp: ºC Gas flow: L/ Nebulizer: psi Capillary: V Agilent Poroshell EC-C. x mm,.7 µm P/N 997- Analytes:. Anandamide (AEA). -arachidonoylglycerol. Impurity. Palmitoylethanolamide (PEA). leoylethanolamide (EA) Poroshell EC-C is less retentive for faster analysis of non-polar compounds Mobile phase: Flow rate: Temperature: ºC Detection: Sample: % CH CN, % H. ml/ nm µl of RRLC checkout sample P/N -9, alkylphenones Agilent Poroshell EC-C, x mm,.7 µm P/N Agilent Poroshell EC-C, x mm,.7 µm P/N

9 A choice of particle sizes to enable you to make the best choice for your method development Easy drop-in replacement for existing μm traditional LC methods This separation of a phenol mix shows the scalability of selectivity going from μm Eclipse Plus C to Poroshell μm and.7 μm columns. It also demonstrates a significant performance improvement in peak capacity. Additionally, the μm column offers imal backpressure increases over existing μm columns, and the.7 μm offers significantly lower backpressure than sub- μm columns. Phenol Mix gradient at. ml/ ZRBAX Eclipse Plus C µm DAD A, Sig=7, Ref=off (CAPACITY\-7-9_--_CSDWS\9.D) Poroshell EC-C µm DAD A, Sig=7, Ref=off (CAPACITY\-7-_--7_CSDWS\PS.D) Poroshell EC-C.7 µm DAD A, Sig=7, Ref=off (CAPACITY\-7-_-9-_CSDWS\7PS.D) Columns: Instrument: All columns. x mm Agilent, pulse damper and mixing column bypassed ZRBAX Eclipse Plus C. µm DAD A, Sig=7, Ref=off (CAPACITY\-7-_--_CSDWS\.D) Mobile phase: A:.% Formic acid B: MeH +.% formic acid Flow rate:. ml/ Temperature: C Detection: nm Sample: steroids Gradient: -% MeH/ Phenol Mix gradient scaled at flows between. and. ml/ Peak Capacity Pressure (bar). Flow Rate (ml/) Flow Rate (ml/) Poroshell EC-C µm Poroshell EC-C.7 µm ZRBAX Eclipse Plus C. µm ZRBAX Eclipse Plus C µm To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell 9

10 ENABLE HIGH-PERFRMANCE SCREENING METHDS WITH NEW PRSHELL HPH-C AND HPH-C 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 (such as acids and bases) can change significantly with varying ph levels, it is becog standard practice to use low, medium, and high ph analyses during method development. New Poroshell HPH-C and HPH-C technologies are made by chemically modifying Poroshell particles using proprietary technology designed to give high ph stability. That means you can use the Poroshell family for all your Fast LC method development needs, regardless of mobile phase ph. Reliable separations for varying ph levels Here, a method utilizing low ph, mid ph, and high ph levels was used to separate 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. 7 ph mm HC NH ph. mm N H HC ph mm NH HC.... Sample:. Procainamide. Caffeine. Acetyl Salicylic Acid. Hexanophenone Deg.. Dipyrimadole. Diltiazem 7. Diflunisal. Hexanophenon

11 Excellent stability at High ph Count on Poroshell HPH chemistries for consistent performance and longevity, even when using high-ph mobile phases. Here, injections of a separation mixture containing acidic, basic, and neutral compounds were performed under extreme ph conditions on an Agilent Poroshell HPH-C and a non-agilent high ph column. Notice the non-agilent column s loss of resolution between the Nortryptyline and Heptanophenone while the Poroshell HPH-C maintains resolution. Column: Agilent Poroshell HPH-C. x mm,.7 µm P/N Injection.... Injection.... Injection.... Injection.... Instrument: Mobile phase: Agilent Infinity Binary LC System A: mm Ammonium Bicarbonate adjusted to ph. in water B: Acetonitrile Column: non-agilent High ph. x mm, µm Flow rate: Gradient:. ml/ Time % B 9. Sample:. Methyl Salicylate. Chlorocinnamic acid. Acetophenone. Quinine. Nortryptyline. Heptanophenone 7. Amitriptyline Injection Injection.... Injection.... Injection....

12 PTIMIZE EVERY SEPARATIN WITH A CHICE F RTHGNAL PHASES Selectivity is the most powerful tool for optimizing HPLC separations. Poroshell EC-C is the best place to start your method development, because of its exceptional flexibility. However, if you are working with challenging analytes, the Poroshell family has many additional chemistries to choose from. For example, our NEW Poroshell PFP columns are engineered with a pentafluorophenyl ligand, which provides an orthogonal separation mechanism with traditional reversed-phase columns. By specifically targeting polar retention mechanisms, 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 and small-molecule pharmaceuticals. Comparative analysis of NSAIDs This separation was completed with four Poroshell chemistries using acetonitrile; each run was only five utes long. nly Poroshell PFP resolved all compounds, although both Poroshell EC-C and Poroshell Phenyl-Hexyl columns eluted the compounds in the same order. The PFP and Bonus-RP columns had very similar elution orders, with the exception of the last two peaks Poroshell PFP 9 9,9., Poroshell EC-C..,7, 9,7, , 9, Poroshell Phenyl-Hexyl Columns: Agilent Poroshell PFP,. x mm,.7 µm P/N Agilent Poroshell EC-C,. x mm,.7 µm P/N Agilent Poroshell Bonus-RP,. x mm,.7 µm P/N Agilent Poroshell Phenyl-Hexyl,. x mm,.7 µm P/N Instrument: Agilent Infinity Binary LC System Mobile phase: A: mm NHHC, ph. B: Acetonitrile Poroshell Bonus-RP Flow rate: ml/ Detection: UV, nm Gradient: Time 7 % rganic. Fluorinated HPLC Phases: Looking Beyond C for Reverse-Phase HPLC M. Przybyciel, LCGC Europe 9() pp 9-,. Sample:. APAP. Phenacetin. Piroxicam. Tolmetin. Ketoprofen. Naproxen 7. Sulindac. Diclofenac 9. Difunisal

13 Analysis of beta blockers: A comparison of Poroshell phases This challenging separation demonstrates how different selectivities produce different results. verall, the Bonus-RP phase delivered the best peak shape and resolution; this was especially true for Naldolol, which appeared as a split peak with the C and Phenyl-Hexyl phases. 7 Poroshell Bonus-RP - - Poroshell Phenyl-Hexyl 7 - Poroshell EC-C 7 Poroshell SB-C 7 - Columns: Agilent Poroshell Bonus-RP,. x mm,.7 µm P/N 97-9 Agilent Poroshell Phenyl-Hexyl,. x mm,.7 µm P/N Agilent Poroshell EC-C,. x mm,.7 µm P/N Agilent Poroshell SB-C,. x mm,.7 µm P/N 77-9 Instrument: Agilent Infinity Binary LC System Flow rate:. ml/ Temperature: C Detection: nm Gradient: % B to % B/ Sample:. Atenolol. Acebutolol. Pindolol. Propranolol. Naldolol 7. Alprenolol. Metoprolol Mobile phase: A: mm NHC, ph. B: MeH To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell

14 Selectivity the most powerful tool for optimizing HPLC separations Poroshell EC-C is the best place to start your method development, because of its exceptional flexibility. However, if you are working with challenging analytes, the Poroshell family has a wide selection of chemistries to choose from. For example, our NEW Poroshell PFP columns are engineered with a pentafluorophenyl ligand, which provides an orthogonal separation mechanism with traditional reversed-phase columns. By specifically targeting polar retention mechanisms, 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 and small-molecule pharmaceuticals. Positional Isomers ( compounds) Water Acetic Acid Acetonitrile Sample:., Dimethoxyphenol., Dimethoxyphenol., Dimethoxyphenol., Difluorophenol., Difluorophenol., Difluorophenol 7., Difluorophenol. Degradation Product, Dimethoxyphenol 9., Dimethylphenol., Dimethylphenol., Dichlorophenol. Chloro methyl phenol. Chloro methyl phenol., Dichlorophenol., Dichlorophenol Time %B Flow Rate ml/ 7 nm. x mm 7 9 Poroshell PFP 7,, 9 7,9 Poroshell EC-C Poroshell Phenyl-Hexyl 7 9 Poroshell EC-C This separation illustrates the benefits of the PFP phase chemistry. Here, positional isomers are analyzed using four different chemistries, with the PFP offering the best resolution of the four chemistries.

15 With the NEW Poroshell µm columns, users can still take advantage of the flexibility of additional phase chemistries. With five chemistries available, users can choose the phase that takes advantage of key analyte interactions, such as the pi-pi interactions shown here with a steroid separation. Isocratic Test Column: Poroshell or PH,. x mm, µm Mobile Phase: % MeCN or MeH % Water w/.% Acetic Acid Sample:. Triamcinalone. Prednisolone. Corticosterone. Estradiol. DES. Dienestrol 7. Deoxycorticosterone Flow Rate:. ml/ Temperature: ºC Detection:, nm,,7 Poroshell µm EC-C 7 7 Poroshell µm Phenyl-Hexyl Steroids separation on both the Poroshell EC-C µm and the Poroshell Phenyl-Hexyl µm chemistries. You can see that from the separation, the better resolution was achieved on the Poroshell Phenyl-Hexyl due to the pi-pi interactions with the analytes and the stationary phase. To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell

16 FAST LC/UHPLC PERFRMANCE FRM A STANDARD HPLC? AGILENT PRSHELL MAKES IT PSSIBLE With Poroshell columns, you can achieve up to -9% or more of the efficiency you would expect from a sub- μm Fast LC/UHPLC column but you can do so at HPLC pressures (below bar). This ability to perform fast separations at low pressures with both the μm and.7 μm column options can dramatically enhance your productivity by allowing you to run more samples in less time using your lab s existing HPLC systems as the following examples illustrate. Plus, you ll be ready to transfer your method seamlessly to an Agilent Infinity Series LC instrument of your choice when you re ready, for even more productivity. UHPLC efficiency with less pressure For this sample of neutral alkylphenones, the Poroshell column delivered >9% of the efficiency attained by the. μm column. Note, too, that the pressure on the Poroshell column is about % of the pressure on the. μm column. Agilent Poroshell EC-C,. x mm,.7 µm P/N N =, Press = bar Mobile phase: % Acetonitrile, % water Flow rate:. ml/ Injection volume: µl Temperature: ºC Detection: DAD Sig =, nm Ref =, nm Sample preparation: RRLC checkout sample P/N -9 spiked w/ µl mg/ml thiourea in water/acetonitrile (:) >9% of the efficiency of. µm Agilent Eclipse Plus C,. x mm,. µm P/N N = 79, Press = bar

17 Choose Agilent Poroshell for high efficiency HPLC In this analysis of soft drink components, the Poroshell column achieved: Agilent Poroshell EC-C,. x mm,.7 µm P/N 997- N =, P = bar >9% of the efficiency of a sub- μm column x the efficiency of the. μm column The pressure on the Poroshell column is below bar, while the pressure on the sub- μm columns is above bar The low backpressure achieved with the methanol mobile phase is especially significant, because methanol generates more pressure than acetonitrile. Column:. x mm,.7 µm Mobile phase: A: %,.% Formic acid B: % Methanol isocratic Flow rate:. ml/ Injection volume: µl Temperature: C Detection: UV, nm Sample:. Saccharin. Caffeine. P-hydroxybenzoic acid Agilent RRHT Eclipse Plus C,. x mm,. µm P/N 999- N =, 7. Aspartame. Dehydroacetic acid. Benzoic acid P = bar Agilent Rapid Resolution Eclipse Plus C,. x mm,. µm P/N 999- N =,9 P = bar Page 7 7 To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell 7

18 Why use a guard column? Simply put, guard columns can save your lab money by extending the life of your analytical column. Installing a less expensive guard column, especially when analyzing dirty samples, prevents damage caused by particulate matter and strongly adsorbed material. As a guide, you should replace your guard column when the plate number, pressure, or resolution changes by more than %; however, you will need to make an exact deteration based on your application. The example here is an accelerated lifetime test with : water/similac with. mg sulfachloropyridazine and sulfamethoxazole. Mobile phase: A:.% Formic acid in water B: Acetonitrile Flow rate:. ml/ Gradient: Hold % B for, ramp to % B in Injection volume: µl Temperature: C Detection: Sig =, nm; Ref = ff Instrument: Agilent Series RRLC Sample preparation: ml Water +. ml similac + ml. mg/ml sulfachloropyridazine and sulfamethoxazole Peak Width () Peak Width () Without a guard column: Analytical column failure after just injections. With a guard column: The guard column fails after injections preserving the analytical column. Injection # Sulfachloropyridazine PW Injection # Replace the analytical column Sulfamethoxazole PW Replace only the less-expensive guard column Fast LC applications stay fast with Agilent Fast Guard columns Agilent Fast Guard columns for UHPLC are rugged and reliable at high pressures, and are fully compatible with Agilent Fast LC and UHPLC columns. They can also be installed without any special tools. Watch our live video demonstration to learn how easy it is to install Agilent Fast Guard columns: agilent.com/chem/poroshell

19 A RUGGED CHICE FR HIGH RESLUTIN LC/MS AND LC/MS/MS Agilent Poroshell columns can make your LC/MS and LC/MS/MS systems work even harder. Their porous outer layer and solid core limit diffusion distance and improve separation speed, while their narrow particle size distribution improves efficiency and resolution. ther advantages include: Quick and efficient resolution of your critical isobaric compounds Better resolution of closely eluting peaks More compounds resolved in a single analysis Improved LC/MS accuracy and identification A standard µm frit which resists plugging with dirty samples Separation of cholesterol and other sterols using Poroshell EC-C columns with LC/MS/MS Note that adequate resolution was obtained, even at the : ratio for cholesterol:lathosterol. This is critical for effective quantitation, because the two compounds have the same molecular weight. Sample:. Calcifediol. Desmosterol. -Cholesten--one. Lathosterol. Cholesterol. Campesterol 7. Stigmasterol. Cholestanol 9. Sitosterol Column: Agilent Poroshell EC-C,. x mm,.7 µm P/N 997- Mobile phase: % Acetonitrile, % methanol Flow rate:. ml/ Injection volume: µl Temperature: C Detection: APCI, positive ion To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell 9

20 D Separation of Vita D /D on Agilent Poroshell EC-C Poroshell provides a very fast LC/MS/MS analysis of Vita D /D in plasma. Isocratic conditions were varied to compare speed of separation with chromatographic resolution. Column: Poroshell EC-C,. x mm,.7 µm P/N Mobile phase: A: H +.% formic acid B: MeH +.% formic acid Flow rate:. ml/ Injection volume: μl Temperature: C Auto sampler temp: C Needle wash: Flush port (::, IPA: MeH:H ) sec Isocratic analysis: A: %, B: % Analysis time: x +ESI MRM Frag=.V CID@. (. -> 9.) VD--.d x Counts vs. Acquisition Time () % MeH +ESI MRM Frag=.V CID@. (. ->.) VD--.d *.7 % MeH -H Vita D -H Vita D Counts vs. Acquisition Time () Rugged performance even after injections This test confirms the outstanding longevity of Poroshell columns, with little performance degradation after injections. Stability is expressed in the consistency of the retention times (%RSD). Analyte %RSD (RT) Analyte %RSD (RT) Analyte %RSD (RT) Morphine.7 Meperidine. Triazolam Codeine. Zolpidem. Naltrexone. Hydrocodone. Fentanyl. Chlordiazepoxide. MDMA. EDDP. Desmethyl diazepam. NorFentanyl. Nitrazepam. Buprenorphine. Heroin. Propoxephine. Cocaethylene. Methyl phenidate. Buprenorphine. -nor-9-carboxy-delta9-thc

21 AGILENT PRSHELL CLUMNS HELP YU INCREASE THE FLEXIBILITY F YUR UHPLC METHDS Because Agilent Poroshell columns have a pressure limit of bar, you can successfully apply them to your UHPLC methods including those that use very long columns, higher flow rates, and viscous solvents. Agilent Poroshell EC-C for fast UHPLC separations This example shows a fast separation using a mobile phase that generates higher pressures. In the top chromatogram, a. mm id column was used, with a flow rate of. ml/ and a pressure below bar making this a typical LC separation. Although the top separation was fast (just under utes), the middle and bottom chromatograms show that you can reduce run times to under utes by increasing the flow rate. These faster analyses will take your pressure to - bar; look to the Agilent Infinity Series flexible upgrade options to help you take advantage of UHPLC capabilities. More viscous solvents like methanol can be used at HPLC or UHPLC pressures. Flow rate =. ml/, P = bar, N BA = 97 Flow rate =.7 ml/, P = bar Flow rate =. ml/, P = 9 bar More viscous solvents like methanol can be used Column: Agilent Poroshell EC-C. x mm,.7 µm P/N 997- Mobile phase: A: %,.% Formic acid B: % Methanol isocratic Flow rate: Varies Injection volume: µl Temperature: ºC Detection: Sig =, nm, Ref = ff Sample:. Saccharin. Caffeine. P-hydroxybenzoic acid. Aspartame. Dehydroacetic acid. Benzoic acid P P P To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell

22 . Agilent Poroshell for HPLC and UHPLC comparison of EPA separation on short and long columns Poroshell columns give you the flexibility to choose longer columns for higher resolution. Here, you can see that as the column gets longer, resolution improves and pressure increases (up to UHPLC pressures for the longest column). Note that resolution is impacted by column length not by the lot of material used in the column proving that Poroshell columns deliver reproducible performance mm length P = bar SN USCFZ7, Lot # B mm length P = bar SN USCFX9, Lot # B mm length P = bar SN USCFW9, Lot # B Balancing column length, resolution, and analysis time are important for any separation. Columns: Agilent Poroshell EC-C,.7 µm Mobile phase: % Methanol, 7% water Flow rate: ml/ Temperature: ºC Agilent Poroshell columns in series deliver the highest efficiency at HPLC and UHPLC pressures Because low backpressure is one of the advantages of Poroshell columns, you can couple several columns in series to achieve the highest separation power per unit time. This enables better separation of complex samples. Peak # Compound Plates k Acetophenone.9 Benzene 99. Touene. Three Agilent Poroshell EC-C,. x mm,.7 µm columns in series for very high efficiency P/N Poroshell Columns in Series. x mm,.7µm Flow rate: ml/ 7. N~ (peak ). Max pressure: bar 7.. Flow rate:. ml/ 7. N~ (peak ). Max pressure: 7 bar Flow rate:. ml/ N~ (peak ). Max pressure: 7 bar 7.. Maximum efficiency at. ml/

23 Fast analysis on an Agilent Poroshell EC-C of common compounds found in analgesics Here, we used a high flow rate to speed up the separation of common analgesic compounds using a Poroshell column., Maximum pressure bar Column: Agilent Poroshell EC-C,. x mm,.7 µm P/N Mobile phase: A: Water +.% formic acid B: Acetonitrile Flow rate:. ml/ Injection volume: µl Temperature: C Detection: DAD, nm Sample:. Acetaophen. Caffeine. -Acetamidophenol. Acetamide. Phenacetin. Sulindac 7. Piroxicam. Tolmetin 9. Ketoprofen. Diflusinal. Diclofenac Keep your LC system running at its best with Agilent A-Line supplies More ways to get more done through: Convenience Simplicity Increased efficiency Learn more: agilent.com/chem/a-line To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell

24 CMPLEX METHD TRANSFERS, MADE SIMPLE Many methods developed on longer µm C columns can be moved to Poroshell columns quickly and easily, especially with the newly available Poroshell µm columns. New changes to the USP regulations are making it easier to transfer conventional methods to newer technologies like Aglient Poroshell. This enables chromatographers to significantly increase throughput and reduce costs. n the following pages, we will show you how five separations, including USP methods, can be repeated on Poroshell columns and can be completed - times faster than the same separations on μm columns. Transfer methods between Agilent Poroshell and ZRBAX for time savings or scalability In this example, a complex method was transferred from a ZRBAX Eclipse Plus C mm, μm column to a mm long Poroshell EC-C column. All conditions were kept the same, except for the gradient time, which was adjusted for the shorter column. As you can see, both separations are the same; however, the bottom chromatogram was generated in just over 7 utes instead of utes for the top chromatogram. An incredible productivity improvement! Keep in d that both separations were run on an older Agilent Series instrument proving that even gradient methods can be transferred while keeping the pressure below bar. bar bar Column: Agilent ZRBAX Eclipse Plus C,. x mm, µm P/N Flow rate: ml/ Sample:. Sulfadiazine. Sulfathiazole. Sulfapyridine. Sulfamerazine. Sulfamethazine.9.. Column: Flow rate:.7.7. Sulfamethazole 7. Sulfamethoxypyridazine. Sulfachloropyridazine 9. Sulfamethoxazole. Sulfadimethoxine...7 Agilent Poroshell EC-C,. x mm,.7 µm P/N ml/ Time %B Time %B. Mobile Phase: A:.% Formic acid in water B:.% Formic acid in acetonitrile

25 Introducing the Agilent Poroshell, μm column expansion This addition provides the Poroshell platform with a scalable solution for chromatographers and method developers. This robust platform extension, initially consisting of the EC-C, EC-C, Phenyl-Hexyl, PFP, and HILIC chemistries enables users to enter the Poroshell family in a very simple manner with the ease of drop-in method replacements. With column pressures that are % less than that of.7 μm Poroshell, and efficiencies nearly double that of traditionally totally porous μm columns, users looking for moderate performance increases can adopt the μm Poroshell columns into their method with simplicity. USP method for Naproxen tablets.x faster analysis on Agilent Poroshell at HPLC pressures This Naproxen separation demonstrates how easy it can be to convert a method to Poroshell columns without changing the flow rate or mobile phase. The st chromatogram shows a USP analysis on an Agilent ZRBAX Eclipse Plus C column, which delivers sharp peaks, three times the needed efficiency, and a resolution of ~. In the nd and rd chromatograms, the Poroshell μm EC-C column ( mm and mm lengths) provides greater efficiency and speed of the original method as easy, drop-in replacements. And because the pressure is only bar for the mm column and 9 bar for the mm column, this isocratic method is an excellent HPLC option. In the th chromatogram, the Poroshell.7 μm EC-C column ( mm length) provides greater efficiency and resolution at nearly x the speed of the original method. The Poroshell EC-C column ( mm length), th chromatogram, still meets the requirements for efficiency and resolution, but is. times faster than the μm System Suitability Method Requirement: N >, Rs >. mau mau x Efficiency x Faster, same backpressure x Faster N =, N =,7 7 Agilent Poroshell EC-C,. x mm (L),.7 µm N =, PN N =,.7 µl injection Rs =. P= bar.x Faster Mobile phase: Flow rate: 7 :9: MeCN:H :acetic acid. ml/ Agilent Poroshell EC-C,. x mm (L),.7 µm PN µl injection Rs = 7. P = bar Agilent Poroshell is an excellent choice for faster methods at HPLC pressures. Sample:. Naproxen. Butyrophenone Agilent ZRBAX Eclipse Plus C,. x mm, µm USP Prescribed Column PN L/dp =, Agilent Poroshell,. x mm [L], µm PN L/dp = 7, Agilent Poroshell,. x mm [L], µm PN L/dp =, Watch our video demonstration to learn how to transfer a Naproxen method to Poroshell columns, and optimize your LC system for the best results. Go to agilent.com/chem/poroshellvideo

26 Fast low pressure analysis Here, a method for analyzing nonnutritive food and beverage additives was transferred from a μm ZRBAX Eclipse Plus C column to an Agilent Poroshell EC-C column, reducing the analysis time from over utes to less than utes. Solvent consumption was reduced by more than % and resolution of the critical pair improved from. to.. Agilent ZRBAX Eclipse Plus, µm, P max = bar Mobile phase: A: mm ammonium acetate, ph. B: Acetonitrile Flow rate: ml/ Temperature: C Gradient: % B at t o, ramp to % B in. Critical pair Rs.79 Agilent Poroshell, P max = bar Mobile phase: A: mm ammonium acetate, ph. B: Acetonitrile Flow rate:. ml/ Temperature: C Gradient: % B at t o, ramp to % B in. Critical pair Rs. Faster analysis of simvastatin on Poroshell Here, a -ute USP method for simvastatin tablets was easily transferred to a Poroshell column, with x faster results. Note that we reduced the column length by 7%, allowing a Poroshell EC-C, 7 mm column to be substituted for a mm long column while still being considered a method adjustment. The Poroshell EC-C phase is similar to other USP L phases, so the results are similar, but faster. Agilent ZRBAX Eclipse Plus C,. x mm, µm P/N Mobile phase: % CH CN, %.9 g/l Na H P, ph Flow rate:. ml/ for µm column. ml/ for.7 µm Poroshell column.7 Temperature: ºC Detection: DAD Sig =, Ref =, nm Agilent Poroshell EC-C,. x 7 mm,.7 µm P/N USP Requirement µm (. ml/).7 µm (. ml/) T R n/a k >..9. N > 99 9 T f <..9.

27 Separation of morphine and metabolites using a Poroshell HILIC column An increasing number of labs are using HILIC early in drug discovery and development for several reasons: To achieve MS compatibility To improve retention of polar compounds and their more polar degradation products To increase LC/MS sensitivity The separation of morphine and metabolites is one example of a fast, efficient HILIC LC/MS method. Here, you can see that these polar compounds are completely resolved in under utes with excellent peak shape and efficiency on the Poroshell HILIC column, where as a reversed-phase method with high aqueous would have limited retention. Agilent Poroshell HILIC,. x mm,.7 µm P/N x TIC SIM Morphine7.d Sample. Normorphine. Morphine. MG. MG Counts (%) vs. Acquisition Time () Mobile phase: A: mm NH HC, ph. B: Acetonitrile mm NH HC, ph : (9:) Flow rate:. ml/ Temperature: C Pressure: 7 to bar System: Agilent 9 Infinity LC and Triple Quadrupole MS Time %B..9 Analysis of vita B and related compounds using a Poroshell HILIC,. x mm,.7 µm column 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 increases LC/MS compatibility and retention. Agilent Poroshell HILIC,. x mm,.7 µm P/N x + TIC SIM Vita.d Counts (%) vs. Acquisition Time () Sample. Aobenzoic acid. Nicotinamide. Riboflavin. Nicotinic acid Mobile phase: Acetonitrile mm NH HC, ph. (9:) Flow rate:.7 ml/ Temperature: C Pressure: bar System: Agilent 9 Infinity LC and Triple Quadrupole MS To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell 7

28 AGILENT PRSHELL CLUMNS: PWERFUL EVERYDAY PRBLEM SLVERS A μm column inlet frit stands up to your dirtiest samples Sub- μm particles offer significant speed and resolution advantages, but are susceptible to clogging with dirty samples because a. µm frit must be used at the column inlet. Poroshell columns solve this problem with a standard μm frit that resists plugging with dirty samples including unfiltered plasma. Sample loading of basic compounds on Poroshell columns is comparable to sub- µm columns Small, non-porous particles have low surface area available for sample interaction, and are limited in their sample loading capability. Poroshell columns, however, are designed with a larger surface area for greater sample loading capacity. In fact, the loading capacity of Poroshell columns is comparable to. μm columns even for the most difficult basic compounds. The peak shape you need for your most accurate results Poroshell columns provide exceptional peak shape especially at ph -7 when compared to other superficially porous columns. Agilent and Series LC systems can easily be optimized for Poroshell columns The inherent properties of Poroshell columns make them ideal for most HPLC and UHPLC instruments, including the new Infinity series LCs. For and series LC systems, all that is needed are or configuration changes (such as flow rate, connector tubing length and id, flow-cell volume, and detector peak-width setting) in order to achieve superior results with lower pressures and higher efficiencies.

29 Agilent Poroshell resists plugging with µm frit Even with dirty samples, such as unfiltered plasma, Poroshell columns show great resistance to plugging. Here, we precipitated the proteins, but did not centrifuge or filter the sample. Even under these conditions, there was no pressure increase, even after injections. Column: Injection volume: Sample: Agilent Poroshell EC-C,. x mm,.7 µm P/N µl injections Precipitated plasma: parts plasma: 7 parts / water-mecn w/.% formic acid with part diflusinal in / water-mecn µg/ml (final concentration diflusinal µg/ml) shaken and allowed to settle utes Not centrifuged/not filtered Diflusinal in plasma Pressure Solvent A: Water w/. % TFA Solvent B: MeCN w/. % TFA Flow Rate: ml/ µl injection Diflusinal in Plasma Injections Time %B Efficiency (N) End Press Plates Instrument: Agilent RRLC (SL) Achieve comparable sample loading to totally porous particles In this example, we loaded nortriptyline (a basic compound) onto several Agilent and competitive columns. Note that the Poroshell.7 μm column has the same loading capacity as the. μm column, and that the. μm column has a broader starting peak width which can compromise resolution. Peak Width Base loading with nortriptyline Agilent Poroshell EC-C,. x mm,.7 µm (USCFX) P/N 997- Agilent ZRBAX Eclipse Plus C,. x mm,. µm (USUYB) P/N Agilent ZRBAX Eclipse Plus C,. x mm,. µm (USUXV) P/N The loads on these columns are typical, proving that Poroshell columns can be used with confidence in basic separations Concentration of Nortriptyline (mg/ml) % mm Na HP buffer, ph., % acetonitrile Temperature: ºC Detection: nm To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell 9

30 Agilent Poroshell columns deliver superior peak shape for better results with basic compounds Here is another basic compound separation, proving how Poroshell columns outperform the competition for challenging analytes. Poroshell Tf =. Competitor S Tf =. Competitor P Tf = Columns: Agilenr Poroshell EC-C,. x mm,.7µm P/N Mobile phase: mm % Na HP, ph 7., % acetonitrile Flow rate:. ml/ Temperature: C Detector: DAD, nm, µl flow cell Sample: µl injection of µg/ml amitriptyline, µg/ml uracil in H /CH CN (9:) k =.9, N=7 Competitor T. x mm N=9, Tf= k =., N= Competitor S. x mm N=, Tf= k =., N= Poroshell EC-C, µm. x mm PN N=, Tf=. Columns: Agilenr Poroshell EC-C,. x mm, µm Mobile phase: mm % K HP / KH P ph 7., % Methanol Flow rate:. ml/ Sample:. UraciI. Propranolol. Butyl Paraben. Dipropylpthalate. Amitriptyline

31 FAST, CNFIDENT SEPARATINS F PRTEINS AND PEPTIDES Agilent Poroshell columns are an excellent choice for separating and characterizing complex bio-molecules, including both intact and digested proteins. Agilent Poroshell columns are the best choice for fast analysis of intact proteins. Agilent Poroshell columns are well suited for peptide mapping, because they provide high resolution with much shorter analysis times than traditional μm columns. For even greater peptide mapping suitability, check out AdvanceBio Peptide Mapping columns, which come pre-tested with a challenging peptide mix to ensure optimal performance for your peptide mapping application. High flow rates with. mm id Poroshell for high resolution and fast separations of proteins Poroshell columns, with their larger pore size and thin shell, are a reliable choice for fast separations of intact proteins. The separation shown here was completed in less than one ute. With their rapid mass transfer of the superficially porous particle, Poroshell columns are the best columns for high efficiency at higher flow rates for extremely rapid separations of proteins. Columns: Mobile phase: Flow rate: Temperature: 7 C Detection: Gradient: Pressure: Agilent Poroshell SB-C. x 7 mm, µm P/N 7-9 A:.% TFA B:.7% TFA in ACN. ml/ UV, nm -% B in. bar.. Time () 7 Sample:. Angiotensin II. Neurotensin. Rnase. Insulin. Lysozyme. Myoglobin 7. Carbonic Anhydrase. valbu Agilent has one of the broadest families of biocolumns available, including AdvanceBio columns to help you advance accuracy and productivity for bioseparations. Learn more at agilent.com/chem/advancebio

32 BioConfirm Molecular Feature Extractor of Stratagene mab trypsin peptide map Using the BioConfirm Molecular Feature Extractor, we can demonstrate % sequence coverage on both the light and heavy chains of the same monoclonal antibody. QTF Instrument Parameters Source ESI positive Gas temperature: ºC Drying gas: L/ Nebulizer: psi Vcap: V Fragmentor: V Skimmer: V ctapole RF: 7 V MS: Hz Mass range: - m/z Reference mass: Acq. mode: Extended dynamic range mode ( GHz) x Counts vs. Acquisition Time () Column: Agilent Poroshell EC-C,. x mm,.7 µm P/N 97- Mobile phase: A: Water,.% formic acid B: ACN,.% formic acid Flow rate:. ml/ Tempurature: C Detection: Gradient: Time %B QTF, ESI positive Shown in table Time %B Insulin analysis: Transfer from a. μm ZRBAX StableBond column to a Poroshell column for increased efficiency Poroshell SB-C column provided double the efficiency of the ZRBAX RRHD SB-C Å due to the larger pore size and more rapid diffusion in the Å pores. The Poroshell columns are ideal for the small protein insulin or other peptides, providing higher efficiency at lower pressure. Agilent ZRBAX SB-C,. x mm,. µm P/N Porcine insulin. A- desamido insulin N = N = 9 Tf =. N =. bar Agilent Poroshell SB-C,. x mm,.7 µm P/N 97-9 N = N = 7 Tf =. Tf =. 9 bar

33 Peptide Map of a Biosimilar EP The top chromatogram shows a peptide map of a highly glycosylated EP from a biosimilar. Note the excellent resolution achieved for small peptide fragments using UV detection. The bottom chromatogram shows the same separation using mass spectroscopy to detere the sequence coverage (%). UV detection is used for comparing peptide maps, while MS is ideal for identifying ao acid substitutions and modifications. So, you can easily confirm protein identity, and identify any post-translational modifications, using the AdvanceBio Peptide Mapping column. Column: Agilent AdvanceBio Peptide Mapping,. x mm,.7 µm, P/N 7-9 Flow Rate:. ml/ Injection: µl (. mg/ml) Temp: ºC Detection: Gradient: nm A, water (.% FA); B, ACN (.% FA), -, -% B; -, -% B; -, -9% B x Cpd :.: +ESI ECC Scan Frag=7.V EPdigestd.d Cpd : Counts vs. Acquisition Time () EP digest, LC/MS TF % sequence coverage achieved using MassHunter Workstation software Agilent AdvanceBio Peptide Mapping columns offer the same Fast LC advantages of Poroshell columns, and are batch-tested with a rigorous peptide mix to ensure suitability and reproducibility. AdvanceBio Peptide Mapping column choices include the new mm length for maximum resolution of the most complex peptide maps. Learn more at agilent.com/chem/advancebio or request Publication No. 99-9EN. To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell

34 Which Fast LC column is best for you? Agilent offers the widest range of Fast LC columns, including Poroshell, ZRBAX Rapid Resolution High Definition (RRHD) columns,. µm (stable to bar) and ZRBAX Rapid Resolution High Throughout (RRHT),. µm (stable to bar). We bond all of these columns with similar stationary phases for assured scalability. With all of these choices, you have flexibility in creating a method to optimize your situation. Your Lab Situation Agilent Recommends Rationale Both UHPLC (+ bar) and HPLC instruments (e.g. Agilent 9 Infinity LC and Infinity LC bar) nly - bar HPLCs Agilent s, Agilent s ( bar) as well as the Infinity LC or Infinity LC ( bar) A mix of UHPLC instruments (Agilent 9 Infinity LC, other + bar instruments) and some HPLC instruments (e.g. LC). Poroshell, μm and.7 μm. ZRBAX RRHD. µm. Poroshell, μm and.7 μm. ZRBAX Eclipse Plus. µm and µm. ZRBAX RRHD. µm. Poroshell,.7 µm Poroshell is an easy column to use on both instrument types. ZRBAX RRHD will help you optimize the capabilities of the 9 Infinity LC for UHPLC. With Poroshell μm and.7 μm columns, you can enhance the performance of older bar instruments, and also get even better performance from newer bar UHPLC instruments. For established methods that you can t transfer, the ZRBAX Eclipse Plus column will provide exceptional peak shape and performance. ZRBAX RRHD can deliver optimum performance on all of these instruments. Poroshell can be used on the bar instruments to optimize their performance. Easy, reliable ph testing, designed for chromatographers Agilent now offers a full line of ph meters and electrodes. Designed for chromatographers, these ph meters offer intuitive user design and exceptional ruggedness for your lab. Agilent CrossLab electrodes are available for non-agilent ph meters. Learn more at agilent.com/chem/agilentph

35 PUSH YUR UHPLC PERFRMANCE T INFINITE LIMITS AND RUN YUR CNVENTINAL METHDS WITH CNFIDENCE Whether you need a workhorse LC system for routine analysis or the most sophisticated, high-resolution LC/MS system, the Agilent Infinity Series has what you re looking for. Together with Poroshell columns, our Infinity Series LC systems deliver ultimate resolution and sensitivity, while helping you boost your separation power per time. They also ensure easy method transferability between systems without redevelopment or revalidation. Infinitely more AFFRDABLE Infinitely more CNFIDENT Infinitely more PWERFUL Learn why Agilent s Infinity Series is infinitely better at agilent.com/chem/infinity Infinity LC Infinity LC 9 Infinity II LC To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell

36 SPECIFICATINS AND RDERING INFRMATIN Agilent Poroshell µm NEW Guard columns for µm NEW Size (mm) EC-C EC-C PFP Phenyl-Hexyl HILIC Size (mm) EC-C. x x x x x x x x x x x x x 7-9. x 7-9. x 7-9 Agilent Poroshell.7 µm HIGH ph Size (mm) EC-C EC-C SB-C SB-C NEW HPH-C NEW HPH-C. x x x x x x x x x x x x x x x

37 Agilent Poroshell.7 µm (continued) Size (mm) Phenyl-Hexyl SB-Aq Bonus-RP HILIC EC-CN NEW PFP. x x x x x x x x x Note: Poroshell columns have a bar/9 psi pressure limit. Agilent Poroshell Fast Guards for UHPLC Size (mm) EC-C EC-C SB-C Phenyl-Hexyl NEW PFP. x x x Agilent Poroshell bonded phase specifications Bonded Phase Pore Size Temp. Limits ph Range Endcapped Carbon Load Surface Area EC-C Å ºC.-. Double % m /g EC-C Å ºC.-. Double % m /g SB-C Å 9 ºC.-. No % m /g SB-C Å ºC.-. No.% m /g HPH-C Å ºC.-. Double Proprietary 9 m /g HPH-C Å ºC.-. Double Proprietary 9 m /g Phenyl-Hexyl Å ºC.-. Double 9% m /g SB-Aq Å ºC.-. No Proprietary m /g Bonus-RP Å ºC.-9. Triple 9.% m /g HILIC Å ºC.-. No N/A m /g EC-CN Å ºC.-. Double.% m /g PFP Å ºC.-. Yes.% m /g Specifications represent typical values only. Unique chemistries increase high-ph stability 7

38 Agilent Poroshell μm Description Size (mm) SB-C SB-C SB-C Extend-C Narrow Bore. x MicroBore. x Capillary. x 7 - Guard Cartridge, /pk. x Guard Hardware Kit MicroBore Guard, /pk. x Note: Poroshell columns have a bar/ psi operating pressure limit. Agilent Poroshell bonded phase specifications Bonded Phase Pore Size Temp. Limits ph Range Endcapped Poroshell SB-C, C, C Å 9 ºC.-. No Poroshell Extend Å C above ph C below ph.-. Yes Specifications represent typical values only. Agilent AdvanceBio Peptide Mapping columns Agilent AdvanceBio RP-mAb columns Description Part Number Description Part Number. x mm,.7 μm 9-9. x mm,.7 μm 9-. x mm,.7 μm 7-9. x mm,.7 μm 7-9. x mm,.7 μm 7-9. mm Fast Guard* 7-9. mm Fast Guard* 7-9. mm Fast Guard* 7-9 * Fast Guards extend column lifetime without slowing down the separation or affecting resolution. C,. x mm,. μm C,. x mm,. μm C,. x mm,. μm C,. x mm,. μm C,. x mm,. μm C,. x 7 mm,. μm C,. x mm,. μm SB-C,. x mm,. μm SB-C,. x mm,. μm SB-C,. x mm,. μm SB-C,. x mm,. μm SB-C,. x mm,. μm SB-C,. x 7 mm,. μm SB-C,. x mm,. μm Diphenyl,. x mm,. μm Diphenyl,. x mm,. μm Diphenyl,. x mm,. μm Diphenyl,. x mm,. μm Diphenyl,. x mm,. μm Diphenyl,. x 7 mm,. μm Diphenyl,. x mm,. μm To learn more about Agilent Poroshell columns, visit agilent.com/chem/poroshell

39 AGILENT CHEMISTRIES: KEEPING YU IN CMMAND F YUR ANALYSES Agilent Poroshell is part of a continuing tradition of performance, expertise and value for small molecule and biomolecule chromatography. We offer alternate selectivities and choices for fast LC separations, including bar ZRBAX RRHD columns,. µm, as well as a range of columns for characterization of biomolecules using size exclusion and ion-exchange. Agilent Bond Elut Silica and Polymeric SPE and Captiva Filtration families of sample prep products offer the widest range of solutions to help you increase throughput and improve the quality of your data. Agilent s meticulous production oversight ensures column and sample prep consistency and performance. With more than years of experience in producing polymers and silica chemistries, our team is committed to continuously developing new advances that make you most productive. 9