synergi Ultra-Performance HPLC The Concept Behind Synergi: Separate the Entire Polarity Spectrum Material Characteristics

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1 Ultra-Performance HPLC Columns The Concept Behind : Separate the Entire Spectrum Much like a carpenter needs a good set of tools, we envisioned analytical chemists would appreciate a unique set of chromatography columns that simplify method development. With the column line, we sought to eliate the guesswork and simplify HPLC column selection, bearing in d the diverse separation scenarios chromatographers might routinely encounter. We engineered column phases to provide improved performance over a set polarity range as well as provide solutions to often difficult to resolve compound mixtures. Whether you are looking to develop a new method or improve an existing one, try it first on a column. Material Characteristics Calculated Packing Particle Pore Pore Surface Carbon Bonded Phase End Material Shape/Size Size Volume Area Load Coverage Capping (μm) (Å) (ml/g) (m /g) % (μmole/m ) Max-RP Spher S Hydro-RP Spher Hydrophilic Polar-RP Spher Hydrophilic Fusion-RP Spher S Max-RP Spher., S Hydro-RP Spher., Hydrophilic Polar-RP Spher., Hydrophilic Fusion-RP Spher., N/A S If does not provide at least an equivalent separation as compared to a competing column of the similar particle size, similar phase and dimensions, send in your comparative data within days and keep the column for FREE. HPLC Mobile Phase Typical Reversed Phase Organic Modified Conditions Polar Selectivity Medium Fusion Hydro Max Non-Polar Polar See p. 6 for information on the Column Selectors. is a trademark of, Inc. 7

2 Maximum Reversed Phase Performance for the Line Max-RP HPLC LC/MS Certified Silanol Activity (ph.) see p. 79 for testing probes. ph Stability.-0.0 Particle Size. µm, µm, and 0 µm Phase Reversed phase C Application For hydrophobic, non-polar compounds over a wide ph range, with little or no MS phase bleed Strength Sharp peak shape for basic compounds at neutral ph C C C with S Endcapping Max-RP C C Improved Polar Selectivity with Reduced Run Times Fusion-RP C8 C8 C8 C8 LC/MS Certified USP:L Silanol Activity (ph.) see p. 79 for testing probes. ph Stability.-0.0 Particle Size. µm, µm, and 0 µm Phase Polar embedded C8 Application For a balanced retention of polar, basic compounds and moderate retention of hydrophobics over a broad ph range Strength Analysis of polar, basic compounds with little or no MS phase bleed C8 with Polar Embedding Fusion-RP Polar Group Hydro-phobic/philic Reversed Phase Retention USP:L Hydro-RP Silanol Activity (ph.) see p. 79 for testing probes. ph Stability.-7. Particle Size. µm, µm, and 0 µm Phase C8 with polar endcapping Application For extreme retention of non-polar and extremely polar alkyl compounds Strength Resolution of highly polar compounds under 00 % aqueous mobile phase conditions C8 with Polar Endcapping Hydro-RP Polar and Aromatic Reversed Phase Selectivity USP:L Polar-RP Silanol Activity (ph.) see p. 79 for testing probes. ph Stability.-7.0 Particle Size. µm, µm, and 0 µm Phase Ether-linked phenyl with polar endcapping Application For extreme retention of polar and aromatic compounds Strength Improved peak shape for acidic and basic analytes and aromatic selectivity with methanol containing mobile phases Ether-Linked Phenyl with Polar Endcapping Polar-RP 8

3 Efficiency. and µm Silica µm Silica Efficiency at Backpressures utilizes a µm 80 Å silica to provide column efficiencies closer to µm columns but at low backpressures similar to µm columns. Ultra-high purity silica (99.99 % metal-free) ensures imal surface metal sites available for chelation and reduces silanol acidity even under neutral mobile phases. With few metal and silanol sites free to bind with sample compounds, peak shapes are sharp. Backpressures Efficiency er Column Backpressure Compared to. µm Competitors App ID 80 App ID 0 App ID Columns: µm Max-RP Waters. µm Symmetry C8 Zorbax. µm SB-C8 Dimensions: 0 x.6 mm Mobile Phase: A = Water with 0. % Formic acid B = Acetonitrile with 0. % Formic acid Gradient: A/B (9:) to A/B (:9) in Flow Rate:.0 ml/ Detection: nm Injection: Sample:. μl. Dexamethasone (. μg). Hydrocortisone (. μg). -α-hydroxyprogesterone (. μg). 7-α-Hydroxyprogesterone (. μg). Progesterone (. μg) µm Max-RP Backpressure=60 Bar Backpressure! Zorbax. µm SB-C8* Backpressure=6 Bar Symmetry. µm C8* Backpressure=8 Bar HPLC *See p. 8 for disclaimer information. Columns: µm Max-RP Luna µm C8 Luna µm C8 Dimensions: 0 x.6 mm Mobile Phase: Water / Acetonitrile (:6) Detection: nm Injection: µl Sample: Naphthalene ( µg) µm C8 µm Max-RP µm C8 Silica Characteristics. µm Silica Rapid Results and Complete Resolution With the increased emphasis on sample throughput; rapid, highresolution techniques are very desirable. Thus we developed a. µm 00 Å ultra-pure silica (99.99 % metal-free) which is packed into the MercuryMS LC/MS cartridge system providing high efficiency, high-throughput screening.. µm Max-RP and Hydro-RP can reduce analysis time by up to 60 % without a loss of performance. Because. µm medias are slurry packed into the MercuryMS cartridges, resolution and peak shapes are equivalent to that typically found only in longer analytical HPLC columns Mercury MS Cartridge System Packed with. µm Max-RP, Hydro-RP, Polar-RP and Fusion-RP For more MercuryMS information, performance and technical specifications, see p. 7. μm 80 Å SILICA Particle Particle Surface Pore Pore Metal Size Size Area Size Volume Content (μm) Distribution (m²/g) (Å) (ml/g) (ppm).00 ± ± 80 ± 0.00 ± 0.0 < 9

4 Max-RP A reversed phase C column If does not provide at least an equivalent separation as compared to a competing column of the similar particle size, similar phase and dimensions, send in your comparative data within days and keep the column for FREE. Maximum Reversed Phase Performance for the Line HPLC Hydrophobic retention similar to a C8 but it s a C % more free silanol coverage than most C8 columns ph stable from. to 0 for sharp peaks for basic and tailing compounds Who says you need a C8 column? The bulky nature of C8 ligands results in relatively low coverage of surface silanols causing peak tailing. Nevertheless, C8 columns have the hydrophobic selectivity chromatographers rely on. To reduce peak tailing and still offer the preferred hydrophobic selectivity, we engineered Max-RP with a C bonded phase. A C ligand is sterically less hindered than a C8 and covers % more of the silica surface, shielding more free silanols. When bonded to our extreme (7 m /g) surface area silica, Max-RP gives the hydrophobic retention and methylene selectivity you would expect from a C8 column, but with sharper peaks, less tailing, and excellent reproducibility. Hydrophobic Retention: Max-RP (C) Performs Like a C8 LC/MS Certified Max-RP C C Silanol Activity (ph.) see p. 79 for testing probes. ph Stability.-0.0 Particle Size. µm, µm, and 0 µm Phase Reversed phase C Application For hydrophobic, non-polar compounds over a wide ph range, with little or no MS phase bleed Strength Sharp peak shape for basic compounds at neutral ph C C C with S Endcapping Max-RP k butylbenzene Max-RP vs. Zorbax. µm XBD C8 Zorbax. µm XDB C XTerra µm MS C8 Zorbax µm SB C8 Nucleosil µm C8 µm Max-RP Zorbax µm XDB C8 Luna µm C8() App ID 9 Columns: XTerra µm MS C8 Zorbax µm SB C8 Nucleosil µm C8 µm Max-RP Zorbax µm XDB C8 Luna µm C8() Dimensions: 0 x.6 mm Mobile Phase: Acetonitrile/ Water (80:0) Flow Rate: ml/ Detection: nm Injection: µl Temperature: Ambient Sample: Butylbenzene App ID 76 µm Max-RP Columns: µm Max-RP Zorbax. µm Eclipse XDB C8 Dimensions: 0 x.6 mm Mobile Phase: A = Water with 0. % Formic acid B = Acetonitrile with 0. % Formic acid Gradient: A/B (9:) to 00 % B in Flow Rate:. ml/ Detection: nm Injection: µl Sample:.Thiourea (0. µg). Codeine ( µg). Chlorphenirae ( µg). Propranolol (6 µg). Desiprae (0. µg) 6. Ibuprofen (6 µg) Sharper Peaks! 0

5 Max-RP (cont'd) Sharper Peaks for Basic Compounds Ionic interactions created by free silanols also can contribute to peak tailing. ph mobile phases (~.) are often employed in order to protonate free silanols. Un-protonated free silanols can lead to poor peak shape for basic drugs. Utilizing a C bonded phase, a % greater bonded phase density is achieved compared to typical C8 bonding, covering more free silanols. Benzylae and phenol are used to probe for active silanol sites at ph.; Max-RP shows lower silanol activity as compared to other C8 columns. Silanol Activity at ph: C vs. C8 Phases alpha of benzylae/phenol Reproducible Performance from ph. to 0 Our bonding and endcapping technology give µm Max- RP stability from ph. (0. % TFA) to 0. This robust ph range ensures that there will be little bleed at low ph s due to bonded phase hydrolysis and that a broad range of mobile phase modifiers can be used without damaging the column. It also allows analysts to use high ph s to overcome basic ionization and to overcome sample solubility issues. ph. App ID 8 Over 8,000 Column Volumes of Mobile Phase! HPLC µm Max-RP Luna µm C8() XTerra µm MS C8 Zorbax µm SB C8 Zorbax µm XDB C8 Nucleosil µm C8 Columns: Dimensions: Mobile Phase: Flow Rate: Detection: Injection: µl Temperature: Ambient Sample: Max-RP vs. Waters XTerra MS XTerra µm MS C8 Zorbax µm SB C8 Nucleosil µm C8 µm Max-RP Zorbax µm XDB C8 Luna µm C8() 0 x.6 mm Methanol/0 mm Potassium phosphate, ph. (0:70) ml/ nm. Benzylae. Phenol ph 0.0 Column: µm Max-RP Dimensions: 0 x.0 mm Part No.: 00A-7-B0 Mobile Phase: A = Water with 0. % TFA B = Acetonitrile with 0. % TFA Gradient: A/B (9:) to A/B (:9) in Flow Rate:.0 ml/ Detection: nm Injection: µl Sample: Precipitated porcine serum (: Acetonitrile:serum) containing:. Oxazepam (0 ng). Temazepam (0 ng). Nordiazepam (0 ng). Diazepam (0 ng) App ID 0896 µm Max-RP Sharper Peaks! Peak Asymmetry =. App ID 8 Over 8,000 Column Volumes of Mobile Phase! XTerra MS µm C8 Peak Asymmetry =.8 Columns: µm Max-RP XTerra MS µm C8 Dimensions: 0 x.6 mm Mobile Phase: 0 mm Potassium phosphate ph 7 / Acetonitrile / Methanol (0::) Flow Rate:. ml/ Detection: 0 nm Injection: µl Sample: Methanol extract from Chlortrimeton Allergy Pills. Chlorphenirae Column: µm Max-RP Dimensions: 0 x.0 mm Part No.: 00A-7-B0 Mobile Phase: Water with 0. % Ammonium hydroxide, ph 0 / Acetonitrile with 0. % Ammonium hydroxide, ph 0 (0:0) Flow Rate: 0. ml/ Detection: nm Injection: µl Sample:. Pyridine. Phenol. Toluene

6 FUSION-RP A polar embedded C8 column If does not provide at least an equivalent separation as compared to a competing column of the similar particle size, similar phase and dimensions, send in your comparative data within days and keep the column for FREE. HPLC Balancing Polar, Basic and Hydrophobic Compound Retention A polar embedded phase for improved separation of polar basic compounds A moderately hydrophobic column for applications requiring moderate retention MS bleed with enhanced selectivity of a polar embedded phase. to 0 ph stability for method flexibility Stable in 00 % aqueous conditions Improved Polar Selectivity with Reduced Run Times Fusion-RP uses a polar embedded and a hydrophobic ligand to achieve improved selectivity. The C8 ligand gives Fusion-RP good hydrophobic retention and selectivity, while the polar embedded group provides enhanced polar retention. This dual-phase selectivity allows balanced polar, acidic, basic and hydrophobic compound retention and resolution. If you are working with mixtures of compounds with polar and non-polar characteristics, and having difficulties finding that perfect mix of selectivity, then you should try Fusion-RP. Fusion-RP LC/MS Certified USP:L C8 Silanol Activity (ph.) see p. 79 for testing probes. ph Stability.-0.0 Particle Size. µm, µm, and 0 µm Phase Polar embedded C8 Application For a balanced retention of polar, basic compounds and moderate retention of hydrophobics over a broad ph range Strength Analysis of polar, basic compounds with little or no MS phase bleed C8 C8 C8 with Polar Embedding Fusion-RP C8 Polar Group Hydrophobic Basic Compounds App ID 80 Longer Polar Retention µm Fusion-RP Less Hydrophobic Retention Balanced Retention! Typical C8 NO-NITROGEN Polar Infused C8 Column By using a non-nitrogen containing polar embedded group, Fusion-RP is able to retain both acidic and basic compounds, with good peak shapes, in comparison to many competing polar embedded columns. The unique polar functionality, in addition to the C8 alkyl ligand, provides a balanced solution to numerous separation needs. Antihistaes App ID 89 Better Selectivity! µm Fusion-RP Waters µm SymmetryShield C8 Columns: Dimensions: Mobile Phase: Flow Rate: Detection: Sample: See p. 8 for ph Stability. µm Fusion-RP Typical C8 0 x.6 mm 0 mm Potassium Phosphate, ph. / Acetonitrile (7:).0 ml/ 0 nm. Maleic acid. Chlorphenirae. Triprolidine. Diphenhydrae Columns: µm Fusion-RP Waters µm SymmetryShield C8 Supelco Discovery µm RP-AmideC6 Dimensions: 0 x.6 mm Mobile Phase: 0 mm Potassium Phosphate, ph. / Methanol (70:0) Flow Rate:.0 ml/ Detection: Sample: & 0 nm. Phenylephrine. Phenylpropanolae. Pseudoephedrine Supelco Discovery µm RP-AmideC6

7 Fusion-RP (cont'd).-0 ph Stable for Rugged Methods The ability of Fusion-RP to operate in an extended ph range of.-0 (under isocratic elution conditions) is the direct result of an exhaustive endcapping procedure, which is highly protective of the silica surface. ph stability is an indication of column ruggedness. ph tested at the extremes (. and 0), for more than 000 column volumes, the results below clearly show how rugged Fusion-RP is. Imagine how well this column will work for your application. ph Stability Test ph ph tr of toluene (.) K of toluene symmetry of toluene symmetry of pyridine N of toluene (plates/column) Column Volumes Extremely LC/MS Column Bleed A careful control of the endcapping process combined with the chemical nature of the polar embedded group results in high phase stability with imal ligand cleavage. The excellent bleed profile compared to a competitor's in the figure below shows Fusion-RP is well suited for LC/MS work. Bleed Profile TIC: Total Ion Current x x0 6 Waters µm SymmetryShield C Dimensions: 0 x.6 mm Mobile Phase: A: 0. % CH COOH in Water B: 0. % CH COOH in Methanol Gradient Rate: 9: (A/B) linear to :9 over 8 hold for Flow Rate: 0. ml/ Detection: Bruker-Daltonics Esquire 000 IT Ion Source: ESI Scan Rate: 000 m/z/s Scan Range: MS Background! µm Fusion-RP HPLC Sulfa Drugs App ID 88 0 ph testing was done under isocratic conditions with phosphate buffer. Formic acid and ammonium formate were also used as test buffers. Enhanced Polar Retention! tr of toluene (.) K of toluene symmetry of toluene symmetry of pyridine N of toluene (plates/column) Column Volumes Waters µm SymmetryShield C µm Fusion-RP 0 00 % Aqueous for Added Method Flexibility Use Fusion-RP for greater polar retention under 00 % aqueous conditions. The polar embedded group allows this phase to be run under 00 % aqueous conditions without loss of retentivity. Unlike typical C8 phases, the pores and the sorbent surface of Fusion-RP stay wet even after many hours of operation in 00 % aqueous mobile phase. This aqueous stability offers greater flexibility in application development. Aqueous Stability App ID At time=0 After hours of 00 % Aqueous Conditions Columns: µm Fusion-RP Waters µm SymmetryShield C8 Dimensions: 0 x.6 mm Mobile Phase: A: 0 mm Potassium Phosphate, ph. B: Methanol Gradient: A/B (9:) for 0. A/B (0:80) in Flow Rate:.0 ml/ Detection: nm Sample:. Sulfanilamide. Sulfathiazole. Sulfamerazine. Sulfamethoxazole. Sulfaquinoxaline Column: µm Fusion-RP Dimensions: 0 x.6 mm Mobile Phase: 0 mm Potassium phosphate buffer ph. Flow Rate:.0 ml/ Detection: nm Injection: µl Sample:. Thiourea. Adenine. Guanosine--monophosphate. Thye

8 Fusion-RP (cont'd) Applications Excellent resolution of nucleic acids, nucleotides, and nucleosides HPLC App ID 86 Fluoropharmaceuticals Column: µm Fusion-RP Dimensions: 0 x.6 mm Part No.: 00F--E0 Mobile Phase: 0 mm Potassium Phosphate, ph./ Acetonitrile (0:0) Flow Rate: ml/ Detection: nm Sample:. Flurbiprofen. Diflunisal App ID 768 Nucleosides Column: µm Fusion-RP Dimensions: 0 x.6 mm Part No.: 00F--E0 Mobile Phase: A: 0 mm Ammonium acetate, ph. B: Acetonitrile Gradient: A/B (9:) to (60:0) in 0 Flow Rate: ml/ Detection: 60 nm Sample:. Cytidine. Uridine. Guanosine. Thye. Adenine App ID Trichothecenes Column: µm Fusion-RP Dimensions: 0 x.0 mm Part No.: 00F--Y0 Mobile Phase: A: 0 mm Ammonium Acetate, ph. B: Acetonitrile Gradient: A/B (90:0) in 0. to (0:80) in Flow Rate: 0. ml/ Detection: Mass Spectrometer (MS) Sample:. T-. -Acetyl-Deoxynivalenol. Nivalenol 6. -Acetyl-Deoxynivalenol. Deoxynivalenol 7. Diacetoxyscirpenol. Fusarenon-X 8. HT- 7 App ID 7 Adenine Family Column: µm Fusion-RP Dimensions: 0 x.6 mm Part No.: 00F--E0 Mobile Phase: A: 0 mm Ammonium acetate, ph. B: Acetonitrile Gradient: A/B (97:) to (60:0) in 0 Flow Rate: ml/ Detection: 60 nm Sample:. ATP. ADP. AMP. Adenine. Adenosine Improve analyte sensitivity and reduce baseline noise with Strata SPE tubes and well plates. See p. for more information. App ID 766 Deoxynucleosides Column: µm Fusion-RP Dimensions: 0 x.6 mm Part No.: 00F--E0 Mobile Phase: A: 0 mm Ammonium acetate, ph. B: Acetonitrile Gradient: A/B (9:) to (60:0) in 0 Flow Rate: ml/ Detection: 60 nm Sample:. '-Deoxycytidine. '-Deoxyuridine. '-Deoxyguanosine. '-Deoxythye. '-Deoxyadenine

9 hydro-rp A polar endcapped C8 column Hydro-phobic/philic Reversed Phase Retention A C8 with enhanced retention of polar compounds Polar resolution enhanced in 00 % aqueous mobile phases Extreme retention of hydrophobic compounds A C8 with Enhanced Retention of Polar Compounds Extremely polar analytes do not retain well nor separate well on conventional C8 columns. Unlike conventional C8 columns, Hydro-RP s C8 bonded phase is endcapped with a proprietary polar group to provide extreme retention of both hydrophobic as well as polar compounds via polar interactions, hydrogen bonding or electrostatic interactions. The high (7 m /g) µm silica surface area combined with a dense bonded phase coverage allows for substantial interaction between the sample analyte and the bonded phase yielding overall a very retentive C8 phase. USP:L Hydro-RP Hydro-RP vs. Zorbax XDB Silanol Activity (ph.) see p. 79 for testing probes. ph Stability.-7. Particle Size. µm, µm, and 0 µm Phase C8 with polar endcapping Application For extreme retention of non-polar and extremely polar alkyl compounds Resolution of highly polar compounds under Strength 00 % buffer mobile phase conditions C8 with Polar Endcapping Hydro-RP HPLC Hydro-RP vs. Luna C8() The polar endcapping of Hydro-RP provides unique selectivity in the separation of basic, polar drugs such as these beta-blockers. Greater retention, polar selectivity! µm Hydro-RP 80 Å Polar Retention! µm Hydro-RP App ID App ID App ID 9 Luna µm C8() 00 Å App ID 66 Agilent Technologies Zorbax µm Eclipse XDB* Dimensions: 0 x.6 mm Mobile Phase: 0 mm Potassium phosphate, ph 7.0 /Methanol (60:0) Flow Rate: ml/ Temperature: C Detection: 0 nm Sample:. Phenylephrine. Phenylpropanolae. Pseudoephedrine. Methylparaben Columns: µm Hydro-RP Zorbax Eclipse XDB µm Dimensions: 0 x.6 mm Mobile Phase: A=0 mm Triethylammonium formate ph 6 B=Acetonitrile with 0 mm Triethylammonium formate Gradient: % B to 6 % B in Flow Rate:. ml/ Temperature: Ambient Detection: 0 nm Injection: µl of beta-blockers mix each at 0.8 µg/ µl Sample:. Atenolol. Pindolol. Nadolol. Acebutolol. Metoprolol 6. Labetalol *See p. 8 for disclaimer information.

10 Hydro-RP (cont'd) Explosives Mix: EPA Method 80 6 Extreme Retention of Hydrophobic Compounds Hydro-RP shows increased hydrophobic retention and polar retention compared to other C8 phases. Increased hydrophobic retention allows for flexibility in fraction manipulation and modification of the organic component in the mobile phase. Enhanced hydrophobic retention allows for separation of compound that differ slightly in hydrophobicity, and results in compounds eluting in higher organic mobile phase, delivering a higher MS sensitivity versus more aqueous eluates. App ID HPLC Hydrophobic Retentivity Compared Column: µm Hydro-RP Dimensions: 0 x.6 mm Part No.: 00G-7-E0 Mobile Phase: Water/Methanol/Acetonitrile (::) Flow Rate: 0.8 ml/ Temperature: C Detection: nm Sample:. HMX. RDX.,,-Trinitrobenzene.,-Dinitrobenzene (Int Std). Tetryl 6.,-Dintrobenzene 7. Nitrobenzene 8.,,6-Trinitrotoluene 9. -Ao-dinitrotoluene 0. -Ao-,6-dinitrotoluene.,6-Dinitrotoluene.,-Dinitrotoluene. -Nitrotoluene. -Nitrotoluene. -Nitrotoluene Dimensions: Mobile Phase: Flow Rate: Temperature: Sample: 0 x.6 mm Acetonitrile/0 mm Potassium phosphate ph 7.0 (6:). ml/ Ambient. Butylbenzene. Amylbenzene The chart was obtained by plotting h y d ro p h o b i c re t e n t i o n ( k f o r butylbenzene vs. methylene selectivity (log k for amylbenzene vs the number of methyl groups) under the stated conditions. A column with high hydrophobicity will better resolve two analytes which subtly differ in their overall hydrophobicity than a column with lower hydrophobic selectivity. Organic Acids App ID 70 Polar retention under 00 % aqueous conditions! Column: µm Hydro-RP Dimensions: 0 x.6 mm Part No.: 00G-7-E0 Mobile Phase: 0 mm Potassium phosphate, ph.9 Flow Rate: 0.7 ml/ Temperature: Ambient Detection: 0 nm Sample:. Oxalic acid. Tartaric acid. Glycolic acid. Formic acid. Pyruvic acid 6. Malonic acid 7. Acetic acid 8. Maleic acid 9. Citric acid 6

11 polar-rp Ether-linked phenyl column with polar endcapping If does not provide at least an equivalent separation as compared to a competing column of the similar particle size, similar phase and dimensions, send in your comparative data within days and keep the column for FREE. Polar and Aromatic Reversed Phase Selectivity Increased retention of highly polar and aromatic compounds Polar resolution enhanced in 00 % buffer mobile phases ly reproducible and stable phenyl phase Polar-RP Increased Retention of ly Polar and Aromatic Compounds Polar-RP is an ether-linked phenyl phase with proprietary hydrophilic endcapping designed specifically to maximize retention and selectivity for polar and aromatic analytes. Aromatic selectivity can be further enhanced by the addition of methanol to the mobile phase. Methanol facilitates the π-π interactions between the aromatic rings of the analyte and the phenyl functional group of Polar-RP. This feature allows for improved polar retention that complements the more conventional C8 column chemistries, as well as provide improved peak shape and an alternative selectivity compared to other polar phases. USP:L Silanol Activity (ph.) see p. 79 for testing probes. ph Stability.-7.0 Particle Size. µm, µm, and 0 µm Phase Ether-linked phenyl with polar endcapping Application For extreme retention of polar and aromatic compounds Strength Improved peak shape for acidic and basic analytes and aromatic selectivity with methanol containing mobile phases Ether-Linked Phenyl with Polar Endcapping Polar-RP HPLC Polar Resolution Compared Improved Resolution and Peak Shape App ID µm Polar-RP Improved Selectivity! Waters µm SymmetryShield C8* Waters µm XTerra RP8* Waters µm Symmetry C8* App ID 090 App ID 0906 Famotidine Peak µm Polar-RP Enhanced Polar Selectivity! Waters µm XTerra RP8* µm Polar-RP Columns: µm Polar-RP Waters µm SymmetryShield C8 Waters µm Symmetry C8 Waters XTerra µm RP8 Dimensions: 0 x.6 mm Mobile Phase: 0 mm Potassium phosphate ph / Methanol (0:0) Flow Rate:.0 ml/ Detection: 0 nm Temperature: Ambient Injection: μl Sample:. Metaproterenol (0. μg). Alprenolol (0. μg). Pindolol (0.6 μg). Propranolol (0.0 μg). Metoprolol (0. μg) 6. Ethylparaben (0. μg) GL Sciences Inertsil µm Phenyl PH-* Zorbax µm SB-Phenyl* Columns: µm Polar-RP Zorbax µm SB-Phenyl Inertsil µm Phenyl PH- Waters XTerra µm RP8 Dimensions: 0 x.6 mm Mobile Phase: 0 mm Potassium phosphate ph 7.0/Acetonitrile (80:0) Flow Rate:.0 ml/ Detection: nm Temperature: C Sample:. Famotidine. Cimetidine. Ranitidine 7

12 Polar-RP (cont'd) Aqueous Stability HPLC Polar Resolution Enhanced in 00 % Buffer Mobile Phases The ether-linkage on the µm Polar-RP phase contributes to not only sharp peak shape for acidic and basic analytes, but also ensures stability under highly aqueous mobile phase conditions. Very polar analytes, like formic acid, typically retain poorly on traditional alkyl-bonded phases. However, using a 00 % buffer mobile phase, the formic acid impurity is easily resolved from acetic acid. Other polar-embedded phases typically use a nitrogen-containing amide linkage or carbamate group; this can interfere with the resolution of highly acidic polar compounds. Since Polar-RP uses an ether-linkage as the polar embedded group, the result is improved peak shape and resolution of the highly acidic polar compound, formic acid. Formic and Acetic Acids App ID 090 µm Polar-RP Better acidic compound resolution in 00 % buffer conditions! Waters XTerra * RP8 µm Column: µm Polar-RP Dimensions: 0 x.0 mm Part No.: 00A-6-B0 Mobile Phase: A = Water with 0. % TFA B = Acetonitrile with 0. % TFA Gradient: A/B (9:) to A/B (:9) in Flow Rate:.0 ml/ Detection: nm Injection: μl Sample: Precipitated porcine serum (: Acetonitrile:serum) containing:. Oxazepam (0 ng). Temazepam (0 ng). Nordiazepam (0 ng). Diazepam (0 ng) Supelco Discovery * RP-AmideC6 µm Columns: µm Polar-RP Waters XTerra RP8 µm Supelco Discovery µm RP-Amide C6 Dimensions: 0 x.6 mm Mobile Phase: 0 mm Potassium phosphate ph./methanol (97:) Flow Rate:.0 ml/ Detection: 0 nm Temperature: C Sample:. Formic acid. Acetic acid Using a Phenyl Phase Phenyl phases can provide separations not achievable on alkyl phases (C8 or C), such as increased retention for polar, aromatic compounds as well as reversals in analyte elution order. The unique selectivity is due to the interactions between the π electrons of the phenyl ring and π electrons from aromatic groups of the analytes.to fully express a phenyl phase s unique aromatic selectivity, methanol should be used as the organic component rather than acetonitrile. It is presumed the π electrons of the CN bond in acetonitrile compete for the phenyl binding sites on the stationary phase, thus limiting the amount of π-π interaction between the stationary phase and the analyte molecules and the separation is based only on hydrophobic retention. *Comparative separations may not be representative of all applications. and Polar-RP are trademarks of, Inc., Luna and are registered trademarks of, Inc. Discovery is a registered trademark of Sigma-Aldrich Biotechnology. has no affiliation with Sigma-Aldrich. Columns used for comparison studies were manufactured by and purchased from Sigma-Aldrich. Nucleosil is a registered trademark of Macherey Nagel. has no affiliation with Macherey Nagel. Columns used for comparison studies were manufactured by and purchased from Macherey Nagel. Inertsil is a registered trademark of GL Sciences Inc.. has no affiliation with GL Sciences Inc.. Columns used for comparison studies were manufactured by and purchased from GL Sciences Inc. SymmetryShield is a trademark of Waters Corp. XTerra and Symmetry are registered trademarks of Waters Corp. has no affiliation with Waters Corp. Columns used for comparison studies were manufactured by and purchased from Waters Corp. Zorbax Bonus-RP is a trademark of Agilent Technologies. Zorbax SB and Zorbax Eclipse XDB are registered trademarks of Agilent Technologies. Columns used for comparison studies were manufactured by and purchased from Agilent Technologies. 8

13 Ordering Information MercuryMS LC/MS Cartridges (mm) 0 x.0 0 x.0 0 x.0 0 x.0 Qty Pricing If does not provide at least an equivalent separation as compared to a competing column of the similar particle size, similar phase and dimensions, send in your comparative data within days and keep the column for FREE.. µm Phase I I I I Hydro-RP 00N-87-BO-CE 00N-87-DO-CE 00M-87-BO-CE 00M-87-DO-CE Max-RP 00N-7-BO-CE 00N-7-DO-CE 00M-7-BO-CE 00M-7-DO-CE Fusion-RP 00N--BO-CE 00N--DO-CE 00M--BO-CE 00M--DO-CE Polar-RP 00N-7-DO-CE 00M-7-BO-CE 00M-7-DO-CE µm Capillary Columns (mm) 0 x x x x x x 0.0 Method Development and Method Validation Kits can be customized for your application. Please call your technical consultant at. Max-RP I 00B-7-AC I 00F-7-AC I 00G-7-AC I 00B-7-AF I 00F-7-AF I 00G-7-AF Hydro-RP 00B-7-AC 00F-7-AC 00G-7-AC 00B-7-AF 00F-7-AF 00G-7-AF µm Microbore and Minibore Columns (mm) SecurityGuard Cartridges 0 x.0 0 x.0 0 x.0 0 x.0 0 x.0 7 x.0 0 x.0 0 x.0 x.0 mm* /0pk Max-RP I 00B-7-A0 I 00F-7-A0 I 00G-7-A0 I 00A-7-B0 I 00B-7-B0 I 00C-7-B0 I 00F-7-B0 I 00G-7-B0 AJ0-607 Hydro-RP 00B-7-A0 00F-7-A0 00G-7-A0 00A-7-B0 00B-7-B0 00C-7-B0 00F-7-B0 00G-7-B0 AJ0-70 Polar-RP 00B-6-A0 00F-6-A0 00A-6-B0 00B-6-B0 00C-6-B0 00F-6-B0 00G-6-B0 AJ0-607 Fusion-RP 00B--A0 00F--A0 00G--A0 00A--B0 00B--B0 00C--B0 00F--B0 00G--B0 AJ0-76 for ID:.0-.0 mm µm Narrow Bore Columns (mm) SecurityGuard Cartridges 0 x.0 0 x.0 7 x.0 0 x.0 0 x.0 x.0 mm* /0pk Max-RP I 00A-7-Y0 I 00B-7-Y0 I 00C-7-Y0 I 00F-7-Y0 I 00G-7-Y0 AJ0-607 Hydro-RP 00A-7-Y0 00B-7-Y0 00C-7-Y0 00F-7-Y0 00G-7-Y0 AJ0-70 Polar-RP 00A-6-Y0 00B-6-Y0 00C-6-Y0 00F-6-Y0 00G-6-Y0 AJ0-607 Fusion-RP 00A--Y0 00B--Y0 00C--Y0 00F--Y0 00G--Y0 AJ0-76 for ID:.0-.0 mm µm Analytical Columns (mm) SecurityGuard Cartridges 0 x.6 0 x.6 7 x.6 0 x.6 0 x.6 x.0 mm* /0pk Max-RP I 00A-7-E0 I 00B-7-E0 I 00C-7-E0 I 00F-7-E0 I 00G-7-E0 AJ0-607 Hydro-RP 00A-7-E0 00B-7-E0 00C-7-E0 00F-7-E0 00G-7-E0 AJ0-7 Polar-RP 00A-6-E0 00B-6-E0 00C-6-E0 00F-6-E0 00G-6-E0 AJ Fusion-RP 00A--E0 00B--E0 00C--E0 00F--E0 00G--E0 AJ0-77 for ID:.-8.0 mm µm Semi-Prep and Preparative Columns (mm) SecurityGuard Cartridges 0 x 0 0 x 0 x. 0 x. 0 x 0 0 x 0 mm x. mm** /pk /ea Max-RP I 00G-7-N0 I 00G-7-AK I 00F-7-P0 I 00G-7-P0 I 00G-7-U0 AJ0-77 I AJ0-77 Hydro-RP 00G-7-N0 00G-7-AK 00F-7-P0 00G-7-P0 00G-7-U0 AJ0-7 AJ0-77 Polar-RP 00G-6-N0 00G-6-AK 00F-6-P0 00G-6-P0 00G-6-U0 AJ0-776 AJ0-77 Fusion-RP 00G--N0 00G--AK 00F--P0 00G--P0 00G--U0 AJ0-78 AJ0-77 for ID: 9-6 mm 8-0 mm 0 µm Analytical, Semi-Prep, Preparative and Pilot Scale Columns (mm) SecurityGuard Cartridges 0 x.6 0 x 0 0 x 0 x. 0 x 0 0 x 0 0 x 0 mm x. mm** /pk /ea Max-RP I 00G-0-E0 I 00G-0-N0 I 00G-0-AK I 00G-0-P0 I 00G-0-U0 I 00G-0-V0 AJ0-77 AJ0-78 Hydro-RP 00G-76-E0 00G-76-N0 00G-76-AK 00G-76-P0 00G-76-U0 00G-76-V0 AJ0-7 AJ0-78 Polar-RP 00G--E0 00G--N0 00G--AK 00G--P0 00G--U0 00G--V0 AJ0-776 AJ0-78 Fusion-RP 00G--E0 00G--N0 00G--AK 00G--P0 00G--U0 00G--V0 AJ0-78 AJ0-78 for ID: 9-6 mm 8-0 mm Axia Packed Columns (mm) 0 x. 00 x. 0 x 0 7 x 0 00 x 0 HPLC µm I I I I I Max-RP 00B-7-PO-AX 00D-7-PO-AX 00B-7-UO-AX 00C-7-UO-AX 00D-7-UO-AX Hydro-RP 00B-7-PO-AX 00D-7-PO-AX 00B-7-UO-AX 00C-7-UO-AX 00D-7-UO-AX Polar-RP 00B-6-PO-AX 00D-6-PO-AX 00B-6-UO-AX 00C-6-UO-AX 00D-6-UO-AX 0 µm Max-RP 00D-0-PO-AX 00B-0-U0-AX 00D-0-UO-AX Hydro-RP 00B-76-UO-AX Polar-RP 00B--UO-AX 0 µm Bulk Packings 00g kg kg Larger quantities of bulk media available upon request. Requires MercuryMS cartridge holder see p. 76 See p. 77 for HST Columns. See p. 8 for Fused Silica Capillary Adapter. See p. 7 for MercuryMS LC/MS Columns. See p. 8 for Axia Packed Columns. Max-RP I 0G-0 I 0K-0 I 0L-0 Hydro-RP 0G-76 0K-76 0L-76 Polar-RP 0G- 0K- 0L- Fusion-RP 0G- 0K- 0L- *SecurityGuard Analytical Cartridges require holder, Part No.: KJ0-8 Semi-prep SecurityGuard Cartridges require holder, Part No.: AJ0-70 **PREP SecurityGuard Cartridges require holder, Part No.: AJ0-8 9

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