Ascentis HPLC Columns

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1 Ascentis HPLC Coumns Eevated HPLC Performance from a Wide Range of Phases Wide range of chromatographic seectivities Enhanced poar compound retention Idea for LC-MS and a modern appications

2 Ascentis: Eevated HPLC Performance We have paced heavy emphasis on optimizing Ascentis phases with reation to the three terms of the resoution equation: efficiency, retention and seectivity. However, our strongest emphasis has been on the most powerfu term, seectivity. Together, Ascentis bonded phases have a wide range of seectivities. It is ikey that one or more Ascentis phase wi accompish any sma moecue HPLC separation. Packed in micro- to preparative hardware dimensions, Ascentis products cover a HPLC appication areas, incuding the most sensitive trace-eve anayses. Ascentis Characteristics The Genera Features of the Ascentis Famiy Incude: High purity, type B siica for inertness, reproducibiity and stabiity Modern bonding processes that optimize bonded phase coverage and maximize stabiity, whie minimizing beed and unwanted secondary interactions Wide seection of bonded phase chemistries and bare siica Phases with enhanced poar compound retention Compatibe with LC-MS and a of today s sensitive instruments and methods Scaabe seectivity from anaytica to preparative High surface area siica for high preparative oading capacity Phase USP Code Key Competitive Feature Modes Primary Uses Page Ascentis C8 L High surface area, inert surface Reversed-phase Sma moecues and peptides 9 Ascentis RP-Amide L60 Phase stabiity, ow beed Reversed-phase Exceent first choice aternative to C8 for routine 0 RP method deveopment. Poar moecues, especiay phenoics and other H-bond donors, acids, bases (uncharged), aniines Ascentis Pheny L Phase stabiity, ow beed Reversed-phase, Ring systems and strong dipoes, p-acids, p-eectron HILIC acceptors, heteroaromatics, nitroaromatics Ascentis ES Cyano L0 Phase stabiity Reversed-phase, Poar compounds, strong dipoes, HILIC, 00% Aqueous tricycic antidepressants Ascentis Siica L High oading capacity, controed and Norma phase Non-poar compounds (in NP mode), highy-poar 6 uniform surface activity (non-aqueous), HILIC compounds (in HILIC mode), nuceosides, amino acids Ascentis C8 L7 High surface area, inert surface Reversed-phase Sma moecues and peptides 8 Discovery HS F* L Orthogona seectivity to C8, ours is Reversed-phase, A eectron and p-eectron donors, bases (charged), 0 we-characterized HILIC, ion-exchange positiona isomers * We have chosen to incude Discovery HS F in this brochure because of its compementary seectivity to the Ascentis phases and its benefits for certain anaytes. Ascentis HPLC coumns represent a continuum of improvement through innovations in HPLC technoogy. sigma-adrich.com/ascentis ordering: (US ony) / 8-9-

3 Deveoping HPLC Methods on Ascentis Seecting An Ascentis Coumn Coumn Screening: The Ascentis & Discovery Method Deveopment Too Kit We recommend every HPLC method deveoper have these five coumns in their arsena. Ascentis C8 Cassic C8 seectivity wi achieve most reversed-phase separations Ascentis RP-Amide For enhanced retention and performance of poar compounds, especiay bases (uncharged) and compounds with H-bond potentia Ascentis Pheny For enhanced retention and performance of poar compounds, especiay ring systems, dipoes, and nitroaromatics Ascentis ES Cyano Extra stabe for ow ph mobie phases due to stericay protected phase. Usefu for poar seectivity in reversed-phase mode. Aso usefu as an aternative to siica in HILIC mode Discovery HS F For enhanced retention and performance of poar compounds, especiay bases (charged) and when the sampe contains a mixture of non-poar and poar compounds Simpy screen these five coumns in your desired mobie phase, using your preferred coumn. Choosing an Ascentis or Discovery Phase Based on Compound Cass and Separation Chaenge or Objective Typicay, Ascentis C8 is the first choice for starting a new method. However, when a C8 doesn t give the desired separation or your sampe contains compounds that are known to be difficut to retain or resove on a C8, consider changing the stationary phase. The range of seectivity provided by Ascentis and Discovery phases makes this easy. The fow chart beow heps guide the seection of Ascentis or Discovery phase based on the particuar compound type or separation chaenge. For more information about each phase, and the other Ascentis phases, pease refer to their dedicated pages in this brochure. Desire to Improve Retention or Resoution on C8 Non-Poar Compounds Mixture of Poar and Non-Poar Compounds Poar Compounds Very Poar Compounds Geometric Isomers Too Much Retention Poor Peak Spacing: Too much retention of non-poar compounds and/or too itte retention of poar compounds Retention Too Short or Inadequate Separation Poor Peak Shape (Basic Compounds) Not Enough RP Retention, LC-MS Not Enough Separation Basic Compounds H-Bonding, Phenoic, Acidic or Hydroxyated Compounds p Eectron- Acceptors Nitroaromatics, Heteroaromatics Strong Dipoes p Eectron- Donors HILIC Mode Reversed- Phase Norma Phase Charged Uncharged Ascentis C8 Discovery Ascentis Ascentis Discovery Ascentis Ascentis Discovery Ascentis HS F RP-Amide Pheny HS F RP-Amide Siica HS F Siica or or or or Ascentis ES Cyano Ascentis ES Cyano Ascentis ES Cyano Ascentis ES Cyano Coumn seection guideines for Ascentis and Discovery phases based on compound cass and separation chaenge or objective. TRADEMARKS: ACE - Advanced Chromatography Technoogies; Ascentis, Discovery, CHIROBIOTIC, CYCLOBOND, CHIRALDEX, CHROMASOLV - Sigma-Adrich Biotechnoogy LP; Fused-Core - Advanced Materias Technoogy, Inc.; Nuceosi - Machery-Nage; Prodigy - Luna; Symmetry, XTerra - Waters Corporation; Zorbax - Agient Technoogies Chromatography Products for Anaysis and Purification

4 Harnessing the Power of Chromatographic Seectivity Chromatographic resoution is a function of coumn efficiency (N), retention (k) and seectivity (a). It is usuay written in the form of the resoution equation: Figure. Affect of Seectivity on Improving Resoution When resoution is potted vs. these three parameters in Figure, it becomes apparent that seectivity has the greatest affect on resoution. The Power to Accompish Difficut Separations One of the most important reasons why seectivity is everaged in HPLC is to resove cosey-euting compounds. A good exampe of this is the need to quantify a compound that eutes in the tai of a more abundant compound, perhaps a ow-eve impurity in the presence of the parent compound, ike shown in the Figure. By atering the stationary phase, in this case going from a C8 to an RP-Amide, the impurity can be euted before the main peak, thereby aowing more sensitive and reiabe quantification. Figure. Impurity Euted on the Downsope of the Main Peak on C8 Difficut to Quantify I C8 Impurity Euted Before the Main Peak on Ascentis RP-Amide Better Quantification Ascentis RP-Amide I The power of chromatographic seectivity is demonstrated in this exampe. The C8 coumn eutes the impurity in the downsope of the major peak, imiting the abiity to detect and quantify the impurity. By using a coumn with different seectivity, in this case an Ascentis RP- Amide, the impurity peak is euted before the main peak. sigma-adrich.com/ascentis technica service: (US and Canada ony) / 8-9-0

5 Key Ascentis Appication Areas Poar Anaytes: Enhanced Retention, Seectivity, and Compatibiity with Highy-Aqueous Mobie Phases Poar compounds are difficut to anayze by traditiona reversed-phase because they ack the high proportion of hydrophobic character necessary for retention. Since most pharmaceuticay- and bioogicay-active compounds are highy poar, this has presented a continua probem in HPLC. From the beginning of our commitment to HPLC innovation, we have focused on bonded phase to enhance poar compound retention. Figure. π-π Interactions (Ascentis Pheny) coumn: Ascentis C8, cm x.6 mm I.D., µm partices (8-U) Ascentis Pheny, cm x.6 mm I.D., µm partices (866-U) mobie phase: 0:0, methano:water fow rate:.0 ml/min. temp.: 0 C det.: UV at nm injection: µl Ascentis Pheny. Uraci. Nitrobenzene.,-Dinitrobenzene.,,-Trinitrobenzene Stationary Phases with Enhanced Poar Compound Retention Compared to C8 Rather than reying soey on dispersive forces to achieve retention, our coumn portfoio contains bonded phases that end additiona retentive character toward anaytes with specific poar functiona groups. Ascentis C8 G0007 Figure. H-bonding (Ascentis RP-Amide) coumn: Ascentis RP-Amide & C8, cm x.6 mm I.D., µm partices mobie phase: 70:0, water with 0.% TFA:acetonitrie fow rate: ml/min. -Hydroxybenzoic Acid temp.: C. Acetysaicyic Acid det.: UV at 0 nm. Benzoic Acid injection: 0 µl. -Hydroxybenzoic Acid. Ethy paraben Ascentis RP-Amide G0007 Ascentis Pheny exhibits strong π-π interactions with the π-acidic nitroaromatics. Addition of each nitro group increases retention. Figure. Ion Exchange (Discovery HS F) coumn: Discovery HS F, cm x.6 mm I.D., µm partices (676-U) conventiona C8, cm x.6 mm I.D., µm partices mobie phase: 0:70, 0 mm ammonium acetate (ph 6.98): CH CN fow rate:.0 ml/min. temp.: C det.: photodiode array injection: µl C8 G0009. Methcathinone. Ephedrine Discovery HS F G0067, C8 G0009 Ascentis RP-Amide exhibits strong retention reative to C8 for phenos and organic acids G0067 Discovery HS F exhibits enhanced retention from ion exchange interactions with ionized anaytes. Chromatography Products for Anaysis and Purification

6 HILIC Mode: Enhanced Retention and High MS-Suitabiity Highy-poar compounds, ike underivatized amino acids, nuceosides and nuceotides, are not we-retained by reversed-phase HPLC. However, under HILIC (hydrophiic interaction chromatography) conditions, they can be retained. HILIC is a variation of norma phase HPLC where the mobie phase contains high percentages of organic modifier. It is aso caed aqueous norma phase or ANP. Under high organic conditions, poar interactions become prominent which can ead to increased retention. Ascentis Pheny, Ascentis ES Cyano, Ascentis Siica and Discovery HS F exhibit HILIC character under highy-organic mobie phases. An added benefit of HILIC mobie phases is the high organic (often >90%) is amenabe to MS detection. Stabiity in Highy-Aqueous Mobie Phases Uness working in HILIC mode, mobie phases for poar compounds are often highy-aqueous, with ony sma percentages of organic modifiers. Under such conditions, C8 phases are not wetted, which causes two probems. First, the bonded phase moecues coaesce resuting in phase coapse and subsequent oss of hydrophobic retention. Second, anaytes have an unpredictabe approach to the siica surface, resuting in irreproducibe and unstabe retention run-to-run and coumn-to-coumn. Ascentis Pheny, Ascentis ES Cyano, Ascentis RP-Amide, and Discovery HS F are competey aqueous-compatibe, and wi not undergo phase coapse even in mobie phases that contain 00% water. Figure 6. Separation of Nuceosides on Ascentis Pheny coumn: Ascentis Pheny, cm x. mm I.D., µm partices (86-U) mobie phase: 0 mm ammonium formate (ph.0 with formic acid) fow rate: 0. ml/min temp.: C det.: UV at 70 nm injection: µl Ascentis Pheny 00% Aqueous- Compatibe 6 7. Cytidine (0. g/l). Deoxycytidine (0. g/l). Uridine (0. g/l). Guanosine (0. g/l). Adenosine (0. g/l) 6. Deoxyguanosine (0. g/l) 7. Deoxyadenosine (0. g/l) 8. Thymidine (0. g/l) 8 Figure 7. Separation of Organic Acids on Ascentis RP-Amide coumn: Ascentis RP-Amide, cm x.6 mm I.D., µm partices (6-U) mobie phase: mm monobasic potassium phosphate (ph.0 with phosphoric acid) fow rate: ml/min. temp.: C det.: UV at 0 nm injection: 0 µl. Dehydroascorbic acid. Ascorbic acid Ascentis RP-Amide 00% Aqueous- Compatibe G00078 G007 Seecting the Right Buffer A partia ist of common buffers and their corresponding usefu ph range is suppied. Perhaps the most common buffer in HPLC is the phosphate ion. Athough, with the growth of LC-MS, voatie buffers such as TFA, acetate, formate, and ammonia are becoming more frequenty used. Remember, the purpose of a buffer in the mobie phase is to inhibit a ph change in the mobie phase after the introduction of a sampe. When deveoping a method, it is important to seect a mobie phase with a fina ph at east one ph unit away from any anaytes pk vaue. As a rue of thumb, one shoud work within a ± ph unit of the buffer pka. Typica buffer concentrations for HPLC tend to be 0-00 miimoar eve. Buffer C Usefu ph Range Trifuoroacetic acid (TFA) 0. <. Phosphate.. -. Formate Acetate Phosphate Ammonia Phosphate sigma-adrich.com/ascentis ordering: (US ony) / 8-9-

7 LC-MS Compatibiity Through Phase Stabiity, Retentivity and Inertness In today s aboratory, HPLC coumns and bonded phases must be compatibe with mass spectrometric detection. Compete MS compatibiity is an important design input for a Ascentis phases. Negigibe Phase Beed Loss of stationary phase can contribute to high background interference in a forms of detection, but it is most notabe in MS detection where phase beed can aso ead to fouing of the instrument and subsequent downtime for ceaning and repair. Modern bonding procedures and an inteigent seection of the Ascentis phase chemistry combine to give a Ascentis phases ow detectabe beed under MS and sensitive UV detection. Amide Chemistry Avoids the Need for TEA and TFA Additives Siano-suppressing mobie phase additives, ike TEA and TFA, are required for good peak shape on traditiona HPLC phases. However, because they suppress the MS signa they shoud be avoided. Ascentis RP-Amide, by virture of the embedded amide group, does not require siano-suppressing additives for good peak shape. Formic acid is a suitabe acidic modifier for use with Ascentis RP-Amide coumns. Ascentis ph Stabiity: Extending the Working ph Range Occasionay, HPLC mobie phases outside the norma ph -7 range are desired to contro sampe stabiity, soubiity or ionization state, or for compatibiity with detection methods. A imitation of most siica-based HPLC phases is their instabiity outside this range where hydroysis of the bonded phase and dissoution of the underying siica can occur. Ascentis coumns have exceent stabiity compared to competitive siica-based coumns. The high bonded phase coverage and proprietary endcapping combine to increase resistance to hydroysis and dissoution. As a resut, Ascentis coumns can be used successfuy between ph.-0 under certain conditions. Note, however, that it is important to avoid storing Ascentis coumns, and any siica-based coumn, in harsh mobie phases. Ascentis Provides Scaabe Separations from Microbore to Preparative Time and precious sampes are wasted during scae-up if the anaytica and preparative coumns do not give the same eution pattern. The high surface area of the underying Ascentis siica provides high oading capacity to purify arger quantities of materia. Additionay, bonded phase and siica chemistry are uniform across,, and 0 µm partice sizes. These features combine to ensure that anaytica separations that are deveoped on Ascentis or µm partices are competey scaabe to preparative separations on Ascentis 0 µm partices and arger coumns. Additionay, separations deveoped on or 0 µm partices can be scaed down for fast anaysis on Ascentis µm partices. Ascentis 0 µm partices in arge coumn dimensions are idea for isoating and purifying mg to gram amounts of compounds for further characterization. Ascentis µm partices in short coumns are idea for rapid anaysis and LC-MS appications. Guideines for Preparing Mobie Phases It shoud be understood that sight variations in ph and buffer concentration coud have a dramatic affect on the chromatographic process; consistent and specific techniques shoud be a reguar practice in the preparation of mobie phases. A common practice is to pace a sufficient amount of pure water into a voumetric fask and add an accurate amount of buffer. The ph of the soution shoud be adjusted, if necessary, and then diute to fina voume of water prior to adding or bending of organic sovents. Then, add a voumetricay measured amount of organic sovent to obtain the fina mobie phase. Thorough bending, degassing, and fitering prior to use is aso recommended. To view a isting of suitabe HPLC and LC-MS additives and sovents, visit sigma-adrich.com/c-ms-sovents Chromatography Products for Anaysis and Purification 7

8 Ascentis C8 The First Choice for Cassic C8 Retention and Seectivity Optimization of siica and bonded phases make Ascentis C8 a true workhorse for the vast majority of HPLC separations. High surface area and phase stabiity give it perfect character for demanding LC-MS and preparative separations. Features: Cassic C8 seectivity High non-poar retentivity Symmetric peak shape Highy reproducibe and stabe Idea for LC-MS Key Appications: Genera reversed phase, hydrophobic and poar compounds Properties: USP code: L Bonded phase description: Octadecy Endcapped: Yes Partice composition: Type B siica ge Partice purity: < ppm metas Partice shape: Spherica Partice size:, and 0 µm Pore size: 00 Å Surface area: 0 m²/g Carbon oad: % ph range (recommended): -8 Extended ph range*:.-0 Characterization of Chromatographic Performance on Ascentis C8 Acidic Ion Exchange Capacity at ph.7 Ion Exchange Capacity at ph 7.6 Hydrogen Bonding Capacity Shape Seectivity Hydrophobic Seectivity Hydrophobic Retention A competitive data obtained from M. R. Euerby, P. Perterson, J. Chromatogr. A, 99 (00) -6; or M. R. Euerby, P. Perterson, W. Campbe, W. Roe, J. Chromatogr. A, (007) 8-. Ascentis data deveoped by Supeco/Sigma-Adrich scientists using the Euerby methodoogy or obtained directy from cited references. Data for Aromatic seectivity is cacuated as atnb/nb/atnb/dnt. Use The cassic reversed-phase coumn, Ascentis C8 is suitabe for any method that specifies a C8-type coumn. Its high surface area gives Ascentis C8 strong hydrophobic retention and high oading capacity for preparative appications. LC-MS Impications Ascentis C8 is ow-beed for cean ESI and APCI traces. The high retentivity means that the mobie phase can contain high eves of organic modifier that are more readiy desovated. Notes Modern C8 coumns have very simiar seectivity, even for basic compounds, because siica quaity and bonding techniques have improved to the point that siano effects are minima. Uness quaity with your current coumn is the issue, the main reasons for evauating different brands of C8 coumns are for improved peak shape and for sight changes in seectivity. * Under certain conditions, the Ascentis famiy can be operated in the extended ph range. For more information, request an eectronic fie of Acid/Base Stabiity of Siica Based on C8, C8, and Amide HPLC Coumns (T0608) 8 sigma-adrich.com/ascentis technica service: (US and Canada ony) / 8-9-0

9 Ascentis C8 is one of the most retentive reversed-phase coumns avaiabe. High retentivity extends the inear range of oading capacity, making Ascentis C8 idea for separations that are or might be used for preparative appications. High retentivity aso means Ascentis C8 can accommodate the highy-organic mobie phases encountered in LC-MS. Ascentis C8: Top of Its Cass in Hydrophobic Retentivity Ascentis C8 Prodigy ODS- Zorbax Extend-C8 Symmetry C8 Luna C8 () Nuceosi C8 ACE C8 XTerra MS C8 Discovery C k Pentybenzene A competitive data obtained from M. R. Euerby, P. Perterson, J. Chromatogr. A, 99 (00) -6; or M. R. Euerby, P. Perterson, W. Campbe, W. Roe, J. Chromatogr. A, (007) 8-. Ascentis data deveoped by Supeco/Sigma-Adrich scientists using the Euerby methodoogy or obtained directy from cited references. Data for Aromatic seectivity is cacuated as atnb/nb/atnb/dnt. Anaysis of basic compounds at neutra ph often gives onger retention compared to acidic mobie phases, but sometimes causes poor peak shape due to siano interactions. The highy-inert surface of Ascentis C8, as with a Ascentis phases, permits anaysis in neutra ph mobie phases. In this exampe, a mix of tricycic antidepressants at ph 7 shows exceent peak shape on Ascentis C8. Figure 8. Symmetrica Peaks for Basic Compounds Indicative of High Surface Deactivation of Ascentis Phases coumn: Ascentis C8, cm x.6 mm I.D., µm partices (6-U) mobie phase: 0:70; mm ammonium phosphate, ph 7.0: methano fow rate:. ml/min. temp.: C det.: UV at nm inj.: 0 µl. Desmethy doxepin. Protriptyine. Desimpramine. Nortriptyine. Doxepin 6. Impramine 7. Amitriptyine 8. Trimpramine G0069 Ascentis C8 provides the seectivity and retention for a range of compounds incuding steroids. Ascentis C8 is a reiabe first choice HPLC coumn that gives symmetric peak shape and exceent retention for even difficut compounds. Figure 9. Estrogenic Steroids on Ascentis C8 coumn: Ascentis C8, cm x.6 mm I.D., µm partices (8-U) mobie phase: :, water:acetonitrie fow rate: ml/min. temp.: C det.: UV at 0 nm inj.: 0 µl. Estrio. a-estrio. b-estrio. Estrone Chromatography Products for Anaysis and Purification 9

10 Ascentis RP-Amide Utra-Low Beed Aky Amide Phase that Rivas C8 as a Generic Scouting Coumn. Exceent Peak Shape and Resoution, Especiay for Poar Compounds or Mixtures of Compound Poarity As pioneers in embedded poar group (EPG) phases for HPLC, Supeco is peased to offer Ascentis RP-Amide, which has a the benefits of enhanced poar compound retention and seectivity, without any of the disadvantages of competitive EPG phases. Features Improved peak shape for bases compared to C8 Different seectivity than C8 or C8 for wide range of poar compounds (especiay acids) Lower beed than competitive EPG phases 00% aqueous compatibe Key Appications Sma, water soube moecues and peptides, H-bond donors, acids, phenos, basic compounds, poar compounds Properties USP code: L60 Bonded phase description: Stabe amide group embedded in an 8-carbon chain Endcapped: Yes Partice composition: Type B siica ge Partice purity: < ppm metas Partice shape: Spherica Partice size:, and 0 µm Pore size: 00 Å Surface area: 0 m²/g Carbon oad: 9.% ph range (recommended): -8 Extended ph range*:.-0 Characterization of Chromatographic Performance on Ascentis RP-Amide Acidic Ion Exchange Capacity at ph.7 Ion Exchange Capacity at ph 7.6 Hydrogen Bonding Capacity Shape Seectivity Hydrophobic Seectivity Hydrophobic Retention A competitive data obtained from M. R. Euerby, P. Perterson, J. Chromatogr. A, 99 (00) -6; or M. R. Euerby, P. Perterson, W. Campbe, W. Roe, J. Chromatogr. A, (007) 8-. Ascentis data deveoped by Supeco/Sigma-Adrich scientists using the Euerby methodoogy or obtained directy from cited references. Data for Aromatic seectivity is cacuated as atnb/nb/atnb/dnt. Use Ascentis RP-Amide can be used for many of the same separations as a C8 whie avoiding some of the disadvantages of C8 such as poor wettabiity in high aqueous mobie phases. In addition, it is much more retentive for those moecues that can interact by hydrophobic interactions and aso by H-bonding with the amide group. Compared to aky-ony phases, Ascentis RP-Amide has enhanced retention and seectivity for phenos, organic acids and other poar soutes due to strong H-bonding between the amide carbony (H-bond acceptor) and H-bond donors, ike phenos and acids. Compared to other EPG phases, ike carbamates, ureas, sufonamides and ethers, Ascentis RP-Amide gives retention comparabe to C8 and C8 for easy coumn comparison without the need to change mobie phase conditions. LC-MS Considerations Unike other amide-based phases, Ascentis RP-Amide uses an amidosiane reagent and a one-step bonding method simiar to C8. Poymeric reagents are aso empoyed to achieve maximum stabiity and ow beed with a modern HPLC detectors. Notes Generay, acids are retained more and bases retained ess on RP-Amide compared to C8 and C8 coumns. Methano can be comparabe in eution strength to acetonitrie when compounds are retained by H-bonding mechanism on the RP-Amide phase. * Under certain conditions, the Ascentis famiy can be operated in the extended ph range. For more information, request an eectronic fie of Acid/Base Stabiity of Siica Based on C8, C8, and Amide HPLC Coumns (T0608) 0 sigma-adrich.com/ascentis ordering: (US ony) / 8-9-

11 Figure 0. Structure of Ascentis RP-Amide G0077 Figure. Enhanced Phenoic Compound Retention via H-bonding on Ascentis RP-Amide coumn: Ascentis RP-Amide, cm x.6 mm I.D., µm partices (6-U) mobie phase: 7:, 0 mm phosphoric acid:acetonitrie fow rate:. ml/min temp.: 0 C OH det.: UV at 70 nm Resorcino injection: µl Because the amide group ies near the siica surface, it is beieved to suppress taiing of basic soutes by eectrostatic shieding (repusion) or by interacting preferentiay with the sianos (H-bonding between the phase and the substrate). The absence of unwanted siano and other secondary interactions gives the Ascentis RP-Amide exceent peak shape for both acids and bases. This exampe of interna deactivation with acetaminophen and pseudoephedrine (Figure ) shows the dramatic effect of changing stationary phase: not ony are the peaks more symmetrica, but eution order is reversed.. Resorcino. Catecho. -Methy resorcino. -Methy catecho.,-dimethy resorcino 6. -Methy catecho 7. -Nitro catecho Catecho Ascentis RP-Amide 6 OH OH G009 OH G0090 Figure. Improved Efficiency, Seectivity and Resoution of Ascentis RP-Amide vs. C8 coumn: Ascentis RP-Amide, cm x.6 mm I.D., µm partices (6-U) Ascentis C8, cm x.6 mm I.D., µm partices (8-U) mobie phase: :8, acetonitrie: mm potassium phosphate fow rate: ml/min. temp.: C det.: UV at nm inj.: 0 µl 7 G009 Ascentis C8. Acetaminophen. Pseudoephedrine C G Ascentis RP-Amide G G00789 An Ascentis RP-Amide coumn is more retentive and seective for phenoic compounds, ike catechos and resorcinos, compared to a C8 and an ether-type poar embedded phase (Figure ). The ether phase does not have the abiity to H-bond with phenoic groups ike the amide group does. Athough comparing an ether phase to a C8 phase may be usefu if ony sighty different seectivity is needed, the most dramatic resuts for acids such as phenos and carboxyic acids wi be obtained with amide-based phases, ike Ascentis RP-Amide. Competitor EPG G Chromatography Products for Anaysis and Purification

12 Ascentis Pheny Utra-ow Beed Pheny Phase with Enhanced Pheny Seectivity Pheny-based reversed-phases were one of the first aternatives to C8 seectivity. Our Ascentis Pheny has been improved to offer exceptiona phase stabiity and enhanced pheny retention. Ascentis Pheny offers versatiity by aso operating in the HILIC mode. Features Low-beed for MS or UV gradient appications due to the use of trifunctiona bonding reagent Outstanding pheny seectivity due to high phase oading and short buty spacer 00% aqueous-compatibe for highy-poar compounds Key Appications Sma, water soube moecues and peptides, π-acceptors, nitroaromatics, poar compounds, dipoes, heterocycics, HILIC mode Properties USP code: L Bonded phase description: Pheny ring with short buty spacer Endcapped: Yes Partice composition: Type B siica ge Partice purity: < ppm metas Partice shape: Spherica Partice size:, and 0 µm Pore size: 00 Å Surface area: 0 m²/g Carbon oad: 9.% ph range (recommended): -8 Extended ph range*:.-0 Characterization of Chromatographic Performance on Ascentis Pheny Acidic Ion Exchange Capacity at ph.7 Ion Exchange Capacity at ph 7.6 Hydrogen Bonding Capacity Shape Seectivity Hydrophobic Seectivity Hydrophobic Retention A competitive data obtained from M. R. Euerby, P. Perterson, J. Chromatogr. A, 99 (00) -6; or M. R. Euerby, P. Perterson, W. Campbe, W. Roe, J. Chromatogr. A, (007) 8-. Ascentis data deveoped by Supeco/Sigma-Adrich scientists using the Euerby methodoogy or obtained directy from cited references. Data for Aromatic seectivity is cacuated as atnb/nb/atnb/dnt. Use Pheny phases are p-basic (eectron donating) and are simiar in overa retention to aky and EPG phases for easy coumn screening. The aternate seectivity of pheny phases is often expained by the p-p interactions avaiabe through the pheny ring. Compounds that appear to exhibit differentia seectivity on the pheny phase incude: hydrophobic bases (TCAs, tetracycines) moderate bases (anesthetics and narcotic anagesics) benzodiazepines some acidic compounds such as ACE inhibitors and quinoine antibiotics nuceosides (e.g. cytidine) nitro, azide and sufony compounds Notes Methano can be a more seective mobie phase component than acetonitrie. Activate HILIC mode by using highy-aqueous (>90%) mobie phases. * Under certain conditions, the Ascentis famiy can be operated in the extended ph range. For more information, request an eectronic fie of Acid/Base Stabiity of Siica Based on C8, C8, and Amide HPLC Coumns (T0608) sigma-adrich.com/ascentis technica service: (US and Canada ony) / 8-9-0

13 Figure. Structure of Ascentis Pheny Si G009 Most commerciay-avaiabe pheny phases actuay show a great dea of C8-ike seectivity, negating their impact on improving the separation. With the highest eve of true pheny character among a phases tested, Ascentis Pheny has exceent aromatic seectivity, making it a true aternative to traditiona C8. The short, buty spacer of Ascentis Pheny does not diute the pheny character as conventiona hexy spacers do. Figure shows the stronger contribution of π-eectron interactions from the pheny ring on Ascentis Pheny compared to the phenyhexy phase aowing it to resove buspirone and trazadone. The pheny and aky seectivity tend to cance each other out on the competitive phenyhexy phase in this case. Aromatic Seectivity - Pheny Ascentis Pheny Luna Pheny Hexy XBridge Pheny Ace Pheny Inertsi Pheny Zorbax SB Pheny Gemini Pheny Hexy Hypersi (BDS) Pheny Betabasic Pheny XTerra Pheny Hypersi Pheny Figure. Better Pheny Seectivity on Ascentis Pheny Compared to Competitive Phases coumn: Ascentis Pheny, cm x.6 mm I.D., µm partices (866-U) mobie phase: 0:60, 0 mm ammonium acetate (ph. with acetic acid):acetonitrie fow rate: ml/min. temp.: C det.: UV at nm injection: µl A competitive data obtained from M. R. Euerby, P. Perterson, J. Chromatogr. A, 99 (00) -6; or M. R. Euerby, P. Perterson, W. Campbe, W. Roe, J. Chromatogr. A, (007) 8-. Ascentis data deveoped by Supeco/Sigma-Adrich scientists using the Euerby methodoogy or obtained directy from cited references. Data for Aromatic seectivity is cacuated as atnb/nb/atnb/dnt. Ascentis Pheny Trazadone. Trazadone. Buspirone G G000, Pheny Hexy type Buspirone G G000 Chromatography Products for Anaysis and Purification

14 Ascentis ES Cyano Extra stabe for ow ph mobie phases due to stericay protected phase Usefu for poar seectivity in the reversed-phase mode, incuding p-p and dipoe-dipoe interacting compounds. Can aso be used in HILIC mode and norma phase chromatography. Features: Enhanced stabiity at ow ph Operates in reversed-phase, HILIC and norma phase modes of chromatography Low MS beed 00% aqueous compatibe Avaiabe as µm and µm partices Key appications: poar compounds, nitroaromatics, tricycic antidepressants, steroids Properties: USP Code: L0 Bonded phase description: diisopropy cyano propy Endcapped: Yes Partice composition: Type B siica ge Partice purity: < ppm metas Partice shape: Spherica Partice size: and µm Pore size: 00 Å Surface area: 0 m /g Carbon oad: 0 % ph range recommended: -8 Use Cyano phases have become very popuar because of their unique seectivity for poar groups and doube bonds. Their potentia for dipoe/dipoe and dipoe/induced-dipoe interaction made them one of the eariest stationary phase functiona groups when aternate seectivity was needed. In the past, stabiity of the cyano phase under reversed-phase and HILIC conditions has been poor. Exposure to aqueous organic mobie phases, acidic ph and eevated temperatures can create gradua retention and seectivity oss that is reportedy due to stationary phase hydroysis. A new Ascentis ES Cyano coumn, based on and µm porous siica substrate, has been deveoped. This new phase compares very favoraby in stabiity to C8, C8, Amide and Pheny. Notes Can be used in reversed, HILIC and norma phase modes It is best to dedicate a specific coumn to one mode of chromatography mentioned above. Methano gives more seectivity than acetonitrie in the reversed phase mode. Cyano phases are used in EPA Method 80 () for the anaysis of exposives and nitroaromatics. Figure. Structure of Ascentis ES Cyano G US EPA Method 80A, Nitroaromatics and Nitramines by High Performance Liquid Chromatography (HPLC) Revision (February 007), obtained from the web site. sigma-adrich.com/ascentis ordering: (US ony) / 8-9-

15 Cyano coumns have been used for the anaysis of tricycic antidepressants for some time. A comparison of the most popuar competitor coumn and the Ascentis ES Cyano phase is shown to the right. Better resoution is seen using the ES Cyano than the competitor coumn for three critica peak pairs. In addition, note the seectivity change of the trimipramine and doxepin (peaks and ) between these two phases under these conditions. Figure 6. Comparison of Tricycic Antidepressants on Ascentis ES Cyano and a Competitor Cyano Phase coumn: as indicated; cm x.6 mm, µm partice mobie phase A: 0 mm potassium phosphate, dibasic (ph 7.0 with phosphoric acid) mobie phase B: acetonitrie mobie phase C: methano mixing proportions: A:B:C, :60: fow rate:.0 ml/min temp.: C det.: nm injection: µl sampe: 00 µg /ml in 70:0, water: methano Ascentis ES Cyano. Trimipramine. Doxepin. Amitriptyine. Imipramine. Nordoxepin 6. Nortriptyine 7. Desipramine 8. Protriptyine G0097 Competitor Cyano G0097 Figure 7. Structures of Tricycic Antidepressants Trimipramine Doxepin Amitriptyin G0097 G0097 G0097 Nortriptyine Nordoxepin Protriptyine G00979 Imipramine G00977 Desipramine G00978 G00976 G00980 Chromatography Products for Anaysis and Purification

16 Ascentis Siica High Surface Area and High Surface Deactivation Combine to Give Ascentis Siica Exceptiona Performance as a Norma Phase, HILIC and Preparative HPLC Materia Besides being the underying support for a Ascentis phases, Ascentis Siica has appications in its own right. Siica is widey used to separate positiona isomers in norma phase mode, and poar compounds in HILIC (aqueous norma phase modes). Siica is aso used in organic synthesis to purify reaction mixtures. In each case, a high purity, controed and uniform surface is necessary to impart the desirabe chromatographic performance. Features High-oading capacity Operates in both norma-phase and HILIC modes Tested in both modes and shipped in ethano, Ascentis Siica is ready to use in either mode Utra-pure, spherica siica Avaiabe in,, and 0 µm Key Appications Sma moecuar weight positiona (geometric) isomers, non-poar compounds (in NP mode), vitamins, steroids, poar compounds (in HILIC mode) Properties USP code: L Bonded phase description: None (surface comprises siano, -Si-OH, and sioxane, -Si-O-Si-, groups) Endcapped: No Partice composition: Type B siica ge Partice purity: < ppm metas Partice shape: Spherica Partice size:, and 0 µm Pore size: 00 Å Surface area: 0 m²/g Carbon oad: 0% ph range (recommended): -6 Use Norma phase and HILIC HPLC modes Preparative chromatography Purification (organic synthesis) LC-MS The cassic use of siica coumns is for norma phase HPLC. The rigid structure of the siica surface, as opposed to the fexibe nature of bonded phases, aows it to distinguish between moecues with different footprints that may have the same hydrophobicity. Geometric isomers and cosey-reated substances, ike the steroids shown in Figure, can be separated on Ascentis Siica under norma phase conditions. Norma phase is aso widey used in preparative chromatography because the mobie phase is more easiy removed by evaporation than the water-containing reversed-phase mobie phases. Figure 8. Ascentis Siica: Norma Phase Separation of Geometric Isomers and Cosey-Reated Compounds coumn: Ascentis Si, cm x.6 mm I.D., µm partices (8-U) mobie phase: 88:, hexane:ethano fow rate:.0 ml/min. temp.: C det.: UV at nm injection: 0 µl sampe: 0 µg/ml in 8:, hexane: -propano Cortisone. Cortisone. Prednisone. Prednisoone G00087 Prednisone Prednisoone G00086 G0008 G sigma-adrich.com/ascentis technica service: (US and Canada ony) / 8-9-0

17 Ascentis Siica is used successfuy in the aqueous norma-phase or HILIC mode. In this mode, water is the strong modifier and the organic is the weak modifier of the mobie phase. Like reversed-phase, HILIC offers the fexibiity of using ph and ionic strength to contro retention. HILIC is idea for very poar compounds and is highy compatibe with LC-MS. Figure 9. Ascentis Siica in HILIC Mode Gives Better Peak Shape and Retention of Basic Drugs than Competitive Siicas coumn: cm x.6 mm I.D., µm partices mobie phase: 0:90, 0.% ammonium acetate in water (ph unadjusted): 0.% ammonium acetate in acetonitrie temp.: C fow rate: ml/min. det.: UV at nm. Heroin. Oxycodone. Codeine. Morphine Ascentis Si Eution order is generay opposite to that obtained under reversedphase conditions. Figure 6 compares Ascentis Siica with two competing siicas, demonstrating better resoution by virtue of exceent peak shape and high retentivity G00089 Inertsi Si G00090 Waters HILIC Si G0009 Poar biomoecues, ike amino acids, nuceotides and nuceosides, typicay require derivatization for their anaysis by reversed phase HPLC. The HILIC mode offered by Ascentis Siica permits the retention and resoution of these compounds without derivatization, eiminating a time-consuming sampe preparation step (Figure 7). Figure 0. Ascentis Siica in HILIC Mode: Amino Acids coumn: Ascentis Si, cm x. mm I.D., µm partices (809-U) mobie phase A: 00 mm ammonium formate (ph.0, with formic acid) mobie phase B: water mobie phase C: acetonitrie temp.: C fow rate: 0. ml/min det.: ESI (+), fu scan inj.: µl sampe: in 0:90, (0 mm ammonium formate/ formic acid, ph.0):acetonitrie Gradient %A %B %C Threonine. Serine. Asparagine. Gutamine. Gutamatic acid G00088 Chromatography Products for Anaysis and Purification 7

18 Ascentis C8 One of The Most Hydrophobic C8 Phases Avaiabe Leveraging the improvements to siica and bonded phase properties that made Ascentis C8 so usefu, its shorter aky chain cousin, Ascentis C8, is aso suitabe for routine HPLC and LC-MS. Features: Seectivity simiar to C8 for non-poar compounds Different seectivity for poar compounds Less hydrophobic than C8, more hydrophobic than other C8 phase Symmetric peak shape Highy reproducibe and stabe Idea for LC-MS Key Appications: Sma, water soube moecues and peptides, ess hydrophobic retention than C8 but comparabe seectivity, LC-MS Properties: USP code: L7 Bonded phase description: Octy Endcapped: Yes Partice composition: Type B siica ge Partice purity: < ppm metas Partice shape: Spherica Partice size: and µm Pore size: 00 Å Surface area: 0 m²/g Carbon oad: % ph range (recommended): -8 Extended ph range*:.-0 Characterization of Chromatographic Performance on Ascentis C8 Acidic Ion Exchange Capacity at ph.7 Ion Exchange Capacity at ph 7.6 Hydrogen Bonding Capacity Shape Seectivity Hydrophobic Seectivity Hydrophobic Retention A competitive data obtained from M. R. Euerby, P. Perterson, J. Chromatogr. A, 99 (00) -6; or M. R. Euerby, P. Perterson, W. Campbe, W. Roe, J. Chromatogr. A, (007) 8-. Ascentis data deveoped by Supeco/Sigma-Adrich scientists using the Euerby methodoogy or obtained directy from cited references. Data for Aromatic seectivity is cacuated as atnb/nb/atnb/dnt. Use Ascentis C8 is suitabe for any method that specifies a C8-type coumn. Athough C8 coumns often show simiar seectivity to C8 coumns, shorter aky chains sometimes show different seectivity toward poar compounds because they can sovate differenty with the mobie phase and interact differenty due to the size and shape of certain moecues. Aso, C8 reagents are smaer than C8 reagents and have improved primary phase coverage, thereby requiring ess end-capping. Ascentis C8 has exceent peak shape and very high phase stabiity. Ascentis C8: Top of Its Cass in Hydrophobic Retentivity Ascentis C8 Symmetry C8 Nuceosi C8 HD RP Seect B C8 Genesis C8 Zorbax SB-C8 HyPurity C8 YMC Basic C8 XTerra MS C8 Discovery C8 0 k Data cited from same source as above. Pentybenzene Among commerciay avaiabe C8 coumns, Ascentis C8 has the highest degree of hydrophobic retention. This permits the use of higher percentages of organic modifier, a benefit to LC-MS users. * Under certain conditions, the Ascentis famiy can be operated in the extended ph range. For more information, request an eectronic fie of Acid/Base Stabiity of Siica Based on C8, C8, and Amide HPLC Coumns (T0608) 8 sigma-adrich.com/ascentis ordering: (US ony) / 8-9-

19 Ascentis C8 is an exceent choice for fast gradient anaysis. Ascentis C8 is typicay more retentive at ow organic composition that C8 and ess retentive at high organic composition. Furthermore, Ascentis C8 has better aqueous compatibiity for gradients that start at 00% aqueous composition. Figure. Fast Gradient Anaysis on Ascentis Coumns coumn: Ascentis C8, cm x.0 mm I.D., µm partices (80-U) mobie phase A: 9:, 0 mm ammonium phosphate:acetonitrie, ph. mobie phase B: 0:60, 0 mm ammonium phosphate:acetonitrie, ph. fow rate:. ml/min temp.: C det.: 0 nm inj.: 0 µl gradient: 0% B to 00% B in min p-touidine. p-nitroaniine. p-nitropheno. p-creso. p-nitrobenzoic acid 6. p-methybenzoic acid 7. p-nitrotouene 8. p-xyene 6 G0077 Ascentis C8 often yieds enhanced retention than Ascentis C8 for sma poar moecues under highy aqueous conditions. Greater retention for Ascentis C8 may be reated to greater wettabiity of Ascentis C8 as compared to Ascentis C8. Figure. Poar Moecues Under Highy Aqueous Conditions coumn: Ascentis C8, cm x.6 mm I.D., µm partices (8-U) mobie phase: :97, methano:water with 0.% TFA fow rate: ml/min. temp.: C det.: UV at 0 nm inj.: 0 µl. L-tartaric. Lactic Acid. Citric Acid. Fumaric Acid. Acryic Acid G0068 Anaysis of bases at neutra ph often yieds enhanced retention over acidic mobie phase but sometimes causes poor peak shape with C8 and C8 coumns due to siano interaction. In this exampe, a mix of tricycic antidepressants at ph 7 shows exceent peak shape on Ascentis C8. Figure. Pharmaceutica Bases at ph 7 coumn: Ascentis C8, cm x.6 mm I.D., µm partices (8-U) mobie phase: 0:70; mm ammonium phosphate, ph 7.0: methano fow rate:. ml/min. temp.: C det.: UV at nm inj.: 0 µl Desmethy Doxepin. Protriptyine. Desimpramine. Nortriptyine. Doxepin 6. Impramine 7. Amitriptyine 8. Trimpramine G0070 Chromatography Products for Anaysis and Purification 9

20 Discovery HS F Unique Reversed-phase Seectivity Compared to C8 and C8 Discovery HS F provides reversed-phase separations that are distincty different from C8 coumns. However, compounds wi generay eute within the same retention time window, making most C8 methods easiy transferabe. Features Unique (orthogona) seectivity compared to C8 and C8 Stabe, ow-beed LC-MS separations Both reversed-phase and HILIC modes Possesses mutipe types of interactions: dispersive, dipoe-dipoe, π-π, charge-transfer Key Appications Sma, water soube moecues and peptides, poar compounds, basic compounds, positiona isomers Properties USP code: L Bonded phase description: Pentafuorophenypropy Endcapped: Yes Partice composition: Type B siica ge Partice purity: <0 ppm metas Partice shape: Spherica Partice size:, and 0 µm Pore size: 0 Å Surface area: 00 m²/g Carbon oad: % ph range (recommended): -8 Characterization of Chromatographic Performance on Discovery HS F Acidic Ion Exchange Capacity at ph.7 Ion Exchange Capacity at ph 7.6 Hydrogen Bonding Capacity Shape Seectivity Hydrophobic Seectivity Hydrophobic Retention A competitive data obtained from M. R. Euerby, P. Perterson, J. Chromatogr. A, 99 (00) -6; or M. R. Euerby, P. Perterson, W. Campbe, W. Roe, J. Chromatogr. A, (007) 8-. Ascentis data deveoped by Supeco/Sigma-Adrich scientists using the Euerby methodoogy or obtained directy from cited references. Data for Aromatic seectivity is cacuated as atnb/nb/atnb/dnt. Notes Compared to a C8: Generay, bases are retained onger on the HS F than on a C8, hydrophobic compounds are retained ess. Increasing the organic content of a C8 separation to 0 percent wi generay provide simiar retention on a HS F. Aso, as a genera rue, soutes with og P o/w vaues ess than. wi be retained onger on HS F compared to a C8. Compared to a pheny phase: Athough aromatic in nature, the pentafuorophenypropy (F) phase does not resembe a pheny phase in retention or seectivity. The F is a strong Lewis acid due to the eectron withdrawing effects of five fuorine groups; the F ring is eectron deficient whereas the pheny ring is eectron rich. Figure. Structure of Discovery HS F G000 0 sigma-adrich.com/ascentis technica service: (US and Canada ony) / 8-9-0

21 Guideines for Transferring a C8 Method to Discovery HS F Generay, bases are onger retained on the HS F than on a C8. Increasing the organic content of a C8 separation to 0 percent wi generay provide simiar retention on a HS F. Resuts with other compounds are highy variabe. However, it is generay true that soutes with ogp o/w vaues ess than. wi be retained onger on HS F compared to a C8. The degree of difference is highy soute dependent. Figure. HS F Provides Exceent Separation - Soutes Are Not Retained on C8 coumn: Discovery HS F, cm x.6 mm I.D., µm partices (676-U) Conventiona C8, cm x.6 mm I.D., µm partices mobie phase: 0:70, 0 mm Ammonium Acetate (ph 6.98): CH CN fow rate:.0 ml/min temp.: C det.: Photodiode Array inj.: µl Discovery HS F G0067,. Methcathinone (00 µg/ml). (+/-) Ephedrine (00 µg/ml) Conventiona C8 No Retention G0067 In Figure, cytidine and reated compounds provide another exampe of the power of HS F to provide unique and vauabe separations compared to a C8. An added benefit of the HS F is its resistance to phase coapse under 00% aqueous conditions. Figure 6. Unique Seectivity of HS F Resoves Compounds Better than C8 coumn: Discovery HS F, cm x.6 mm I.D., µm partices (676-U) Conventiona C8, cm x.6 mm I.D., µm partices mobie phase: 0 mm KH PO, ph.0 with H PO, (C8 separation has % CH CN) fow rate: ml/min temp.: 0 C det.: UV at 80 nm inj.: 0 µl, each compound 00 µg/ml. Cytidine. Cytosine. -Deoxycytidine Discovery HS F 0 6 G009 Conventiona C8 0 6 G009 Chromatography Products for Anaysis and Purification

22 Extreme Performance on Any LC System The demand for increased sampe throughput and speed of resuts has driven HPLC users to search for breakthroughs in HPLC instrument and coumn technoogy. Athough improvements have been reaized, setbacks have been encountered. Reductions in coumn ruggedness, costy repacements of existing instrumentation, and difficuties in transferring methods to new systems have often made these past improvements unappeaing to anaysts. Ascentis Express has changed a of that. Ascentis Express with Fused-Core Partice Technoogy provides the utimate soution for today s separation demands - high speed and high efficiency with ow backpressure. By simpy changing to Ascentis Express Coumns, sampe throughput can be improved by 00%! No onger wi you have to make changes to: sampe prep fow rate system pressure And no new instrumentation is required! For more information on this exciting new technoogy, visit our website: sigma-adrich.com/express Do More Work in Less Time without Changing your Method With identica conditions, the Ascentis Express C8 coumn performs X as many separations as a standard C8 coumn in ess time. CONDITIONS Theoretica Standard C8 Ascentis C8 Sampe Prep Fow Rate System Pressure HPLC System Pates Throughput Throughput SAME SAME SAME SAME SAME X X C8, cm x.6 mm I.D., μm partices mobie phase: 6:, water:acetonitrie fow rate: ml/min. temp.: ambient det.: UV at nm. Oxazepam: FW = 86. Aprazoam: FW =. Coazepam: FW =. N-desmethydiazepam: FW = 70. Diazepam: FW = 8 X Ascentis Express C8, 0 cm x.6 mm I.D.,.7 μm partices (87-U) X X X X sigma-adrich.com/ascentis ordering: (US ony) / 8-9-

23 Ordering Information Partice ID Length Ascentis Ascentis Ascentis Ascentis Ascentis Ascentis Discovery Size (mm) (cm) C8 RP-Amide Pheny ES Cyano Siica C8 HS F µm µm 0 µm 8-U 609-U inquire inquire inquire 8-U inquire 0 86-U 689-U 8600-U inquire 80-U 8-U inquire 86-U 666-U 860-U inquire 8-U 86-U inquire. 8-U 6-U inquire inquire inquire 8-U inquire. 8-U 6-U 860-U inquire 8-U 8-U 670-U. 800-U 600-U 860-U 7708-U 800-U 800-U 6700-U U 60-U 860-U 7709-U 80-U 80-U 670-U. 80-U 60-U 860-U 770-U 80-U 80-U 670-U 8-U 6-U inquire inquire inquire 8-U inquire 80-U 60-U 8606-U inquire 8-U 80-U 670-U 807-U 6-U inquire inquire inquire 80-U inquire U 6-U 8607-U inquire 80-U 80-U 678-U.6 8-U 66-U inquire inquire inquire 86-U inquire.6 86-U 67-U inquire inquire inquire 87-U 6709-U.6 80-U 60-U 8608-U 77-U 80-U 806-U 670-U U 6-U 8609-U 77-U 80-U 807-U 670-U.6 8-U 6-U 860-U inquire 806-U 808-U 6707-U. 868-U 69-U inquire inquire inquire 89-U inquire. 87-U 6-U inquire inquire inquire 80-U inquire. 80-U 60-U 86-U 7700-U 807-U 80-U 6708-U U 60-U 86-U 770-U 808-U 89-U 670-U. 80-U 60-U 86-U 770-U 809-U 8-U 67-U. 80-U 606-U 86-U inquire 80-U 8-U 67-U 88-U 6-U inquire inquire inquire 8-U inquire 869-U 69-U inquire inquire inquire 80-U inquire 89-U 6-U inquire inquire inquire 8-U inquire.6 80-U 6-U inquire inquire inquire 8-U inquire.6 8-U 66-U inquire inquire inquire 8-U inquire.6 8-U 6-U 86-U 770-U 8-U 8-U 67-U.6 0 inquire 68-U inquire 770-U inquire inquire 67-U.6 8-U 6-U 866-U 7706-U 8-U 8-U 676-U.6 8-U 6-U 867-U 7707-U 8-U 8-U 677-U 0 80-U 60-U inquire inquire inquire inquire 678-U U 6-U inquire inquire inquire inquire 677-U 0 8-U 6-U inquire inquire inquire inquire 679-U 0 8-U 6-U 868-U inquire 8-U inquire 670-U. 8-U 6-U inquire inquire inquire inquire inquire. 87-U 68-U 869-U inquire 8-U inquire 67-U.6 80-U 6-U inquire inquire inquire inquire inquire.6 8-U 6-U inquire inquire 8-U inquire inquire 0 8-U 6-U inquire inquire inquire inquire inquire U 6-U inquire inquire inquire inquire inquire 0 8-U 66-U inquire inquire inquire inquire inquire 0 8-U 67-U inquire inquire 86-U inquire inquire. 86-U 68-U inquire inquire inquire inquire inquire U 69-U inquire inquire inquire inquire inquire. 88-U 660-U inquire inquire inquire inquire 678-U. 89-U 66-U inquire inquire 87-U inquire 679-U ID Length Partice Size Ascentis Ascentis Ascentis Ascentis Ascentis Discovery (mm) (cm) (µm) C8 RP-Amide Pheny Siica C8 HS F Ascentis Supeguard Cartridges Kit. 876-U inquire inquire inquire inquire 677-U Pack of. 877-U inquire inquire inquire inquire 6770-U Pack of. 870-U 67-U inquire inquire inquire 677-U Kit. 87-U 67-U inquire inquire inquire 677-U Pack of 87-U 67-U inquire inquire inquire inquire Kit 87-U 67-U inquire inquire inquire inquire Kit 878-U inquire inquire inquire inquire 677-U Pack of 879-U inquire inquire inquire inquire 677-U Pack of 87-U 670-U 860-U 88-U 86-U 6776-U Kit 87-U 67-U 86-U 89-U 87-U 6777-U Kits incude one cartridge, stand aone hoder, a piece of tubing, nuts and ferrues. Ascentis Vaidation Packs U 69-U 869-U inquire inquire inquire.6 89-U 69-U 8696-U inquire inquire inquire Vaidation Packs incude coumns, each from a different bond ot. Chromatography Products for Anaysis and Purification

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