Elevated HPLC Performance

HPLC Columns from Supelco

RP-Amide

C18

sigma-aldrich.com/supelco 2 Supelco – A Pioneer in Chromatography

upelco’s 30-year history in pioneering Fluorinated phases are another area of pioneering Schromatography products has led to research for Supelco. Discovery® HS F5 is a market worldwide recognition of its products. leader providing reversed phase as well as aqueous normal phase (ANP) or hydrophilic interaction (HILIC) Supelco introduced SUPELCOSIL™ LC-18-DB, the ﬁ rst separations. base deactivated HPLC column, in 1982. It provided Supelco, along with Sigma-Aldrich, Fluka, and improved peak shape for basic compounds that Riedel-de Haën, provides thousands of chromatogra- traditionally tailed with early generation columns. phy related products through a worldwide R&D, Supelco also innovated with the amide-type embed- manufacturing, distribution and sales operation. This ded polar group (EPG) column, Suplex™ pKb-100, and includes sales and distribution in over 35 countries and has been a leader in EPG bonding chemistry. R&D and production on four continents.

Sigma-Aldrich... The World’s Leading Supplier of Analytical Products sigma-aldrich.com/supelco

ASCENTIS™ HPLC COLUMNS 3 The Ascentis Family of HPLC Columns

The Ascentis family of columns is the fourth generation provides quality oversight into all aspects of the tion of HPLC column technology from Supelco scien- manufacturing process leading to a product that tists. Ascentis columns are bonded on high purity, consistently meets exacting speciﬁ cations. 100 angstrom silica including 3, 5, and 10 micron Supelco’s bonding chemistry advances have particle size. Columns are designed for small molecule achieved: applications and are scalable from micro columns ● Improved stability for extremely long column life (1.0 mm I.D.) to preparative dimensions (50 mm I.D.). ● Better reproducibility for easy method validation The family includes C18, C8 and an embedded polar ● group phase, RP-Amide. Low bleed for sensitive LC-MS applications ● Columns are manufactured at our ISO 9001 regis- Excellent selectivity and increased retention tered facility in Bellefonte, PA. This ISO 9001 registra- for polar compounds

Ascentis Separates the Entire Polarity Spectrum / 814-359-3041 Ascentis HPLC Columns (US and Canada only)

Column Speciﬁ cations C18 C8 RP-Amide USP Code L1 L7 L60 Bonded Phase Octadecyl Octyl Amido embedded Endcapped Yes Yes Yes Particle Shape Spherical Spherical Spherical Particle Purity <5 ppm metals <5 ppm metals <5 ppm metals Particle Size 3,5,10 µm 3,5 µm 3,5,10 µm Pore Size 100 Å 100 Å 100 Å Surface Area 450 m2/g 450 m2/g 450 m2/g Carbon Load 25% 15% 19.5% pH Range 2 - 8 2 - 8 2 - 8 / 814-359-3441 technical service: 800-359-3041 (US only) ordering: 800-247-6628 ordering:

Elevated HPLC Performance 4 Extended Column Life Save time and money

One of the most important parameters surface area of the silica help to create a more consistent in choosing a column is lifetime. and well-packed column that minimizes shifting of the Columns that last longer save time and money. Column column bed during use. This reduces the chance of voiding. failure often occurs because of hydrolytic instability of the Supelco scientists have performed extensive column stationary phase and silica as well as mechanical instability lifetime studies under conditions employed in most HPLC of the column packing (voiding). Ascentis columns are laboratories. Studies included the use of pH modiﬁ ers such designed to last beyond competitive columns. as TFA, phosphate, and ammonia coupled with both The high phase coverage of all Ascentis products plus acetonitrile and methanol conditions. A range of tempera- proprietary endcapping create a phase that is more stable tures was explored across a variety of probe analytes toward hydrolytic attack. Supelco’s narrow speciﬁ cations on including neutrals, acids, bases, and pharmaceuticals.

Extended Column Life Even Under Harsh pH 1.5 Conditions

12 Extended Column Life 10 Toluene

k’ 6

4 2,4-dichlorophenoxyacetic acid

0 0 2000 4000 6000 8000 Number of Injections* *Equivalent to 80,000 column volumes

Quality Control Application First Run Symmetry and selectivity remain unchanged on 1 this sensitive application even after 80 days▲ of pH 5 4 1.5 conditions. (▲100 injections a day) 2

column: Ascentis C18, 5 cm x 3.0 mm I.D., 5 µm particles (581329-U) mobile phase: 60:40, methanol:100mM ammonium acetate, pH 6.8 ﬂ ow rate: 0.43 mL/min. G003379 temp.: 25 °C 3 det.: 230 nm inj.: 5 µL After 80 Days 1. Uracil 2. Hydroquinidine 3. Quinidine 1 4. Fluoxetine 5 4 5. Diphenhydramine 2

G003380 sigma-aldrich.com/supelco

ASCENTIS™ HPLC COLUMNS 5 Highly Reproducible HPLC Columns Minimize troubleshooting

Reproducibility is imperative to any Bonding procedures are transferred to production through application in any laboratory. a manufacturing transfer validation procedure. Each bonded lot is analyzed by QC and must pass bond lot specifi cations A novel column may be interesting, but if the retentivity before a single column is packed. Once the bond lot meets and selectivity are not consistent, it has little value in a specifi cations, each individual column must meet QC validated method. Supelco has designed reproducibility into specifi cations prior to being tagged with the Ascentis label. the Ascentis family starting with tight specifi cations on the Detailed production and QC documents are maintained for surface area, pore size, and particle size of the silica. every column that receives the Ascentis label.

Carbon Load on Ascentis C18 Since Inception Columns

30.0

NOTE: 1.0% RSD for carbon load. 28.0 Highly Reproducible Highly Reproducible / 814-359-3041

26.0

% Carbon 24.0 (US and Canada only)

22.0

20.0 Carbon load data obtained from 3, 5, and 10 micron bond lots.

Ascentis RP-Amide Reproducibility - Aromatic Compounds

column: Ascentis RP-Amide, 15 cm x 4.6 mm I.D., 5 µm particles (565324-U) mobile phase: 30:70, water:acetonitrile NOTE: Less than 1.0% ﬂ ow rate: 1.0 mL/min RSD for retention times. temp.: 35 °C det.: UV, 254 nm inj.: 2 µL

1. Butylbenzene 2. o-Terphenyl 3. Triphenylene 4. Amylbenzene

Butylbenzene Triphenylene / 814-359-3441 technical service: 800-359-3041

BL Rt USP Tail Factor N Rt USP Tail Factor N 5403A 8.66 0.950 13657 13.03 0.959 13746

5422A 8.69 0.923 12527 12.88 0.919 12594 (US only) 5433 8.76 1.000 14101 13.06 1.010 14350

Avg. 8.700 0.957 13428 12.980 0.959 13563 St. Dev. 0.051 0.039 811 0.102 0.040 892 % RSD 0.580 4.070 6.04 0.780 4.220 6.57 Bond Lot 5433 Bond Lot 5422A Bond Lot 5403A

G003390 ordering: 800-247-6628 ordering:

Elevated HPLC Performance 6 Ideal for LC-MS Short, Fast Columns

Multiplying Productivity with lower solvent consumption. Ascentis columns work well for Elevated Performance fast LC because they are rugged and more retentive than Imagine facing more and more samples each month other reversed phase columns. Greater reversed phase without an increase in chemists or instruments. The solution retention allows more organic solvent that lowers viscosity to this dilemma is here today with Ascentis fast HPLC and pressure. Another potential beneﬁ t of operating with columns. Fast LC performs rapid analysis using short (2-10 higher organic mobile phases is increased performance from cm) columns operating in the 1-4 mL/min range. Fast LC has evaporative detectors such as MS. Ascentis fast LC columns many beneﬁ ts including faster analysis, reduced method are available with all three phases (C18, C8, and RP-Amide)

LC-MS development time, increased mass spec sensitivity, and with column lengths down to 2 cm.

Fast Gradient Analysis on Ascentis Columns A comparison of all three Ascentis phases under fast 8 Ascentis RP-Amide 3 gradient conditions is shown. A set of aromatics with a 5 variety of functional groups was utilized. The interactions 4 7 NOTE: Elution between the anilines, phenols, and carboxylic acids and the order is reversed Ascentis RP-Amide give different selectivity as compared to for peaks 4 and 5. the Ascentis C18 and C8. When comparing the Ascentis C18 6 and C8, the Ascentis C8 is more retentive at lower organic 1 2 while the Ascentis C18 is more retentive at higher organic.

column: Ascentis RP-Amide, 3 cm x 3.0 mm I.D., 5 µm particles Ascentis C18, 3 cm x 3.0 mm I.D., 5 µm particles Ascentis C8, 3 cm x 3.0 mm I.D., 5 µm particles G003375 mobile phase A: 95:5, 10 mM ammonium phosphate:acetonitrile, pH 2.5 mobile phase B: 40:60, 10 mM ammonium phosphate:acetonitrile, pH 2.5 4 ﬂ ow rate: 3.5 mL/min Ascentis C18 5 8 temp.: 35 °C 3 det.: 220 nm inj.: 10 µL 7 gradient: 0% B to 100% B in 4 min.

1. p-Toluidine 2. p-Nitroaniline 3. p-Nitrophenol 6 4. p-Cresol 5. p-Nitrobenzoic acid 1 2 6. p-Methylbenzoic acid 7. p-Nitrotoluene 8. p-Xylene

G003376

4 Ascentis C8 5 8 3

1 2

G003377 sigma-aldrich.com/supelco

ASCENTIS™ HPLC COLUMNS 7

Fast Gradient Analysis on an Ascentis RP-Amide Column with MS Detection

A column lifetime study was performed using a fast 2 5 gradient to separate caffeine (major component) and catechins from green tea extract. Even after 500 injections, 1st Injection all seven peaks are still resolved in under two minutes. 3 7

6 column: Ascentis RP-Amide, 2 cm x 4.6 mm I.D., 3 µm particles (565316-U) mobile phase A: 10 mM ammonium formate, pH 3.0 with concentrated formic acid mobile phase B: methanol 4 ﬂ ow rate: 1 mL/min., split to the mass spectrometer temp.: 35 °C det.: MS, ESI (-) and (+) in selected ion recording (SIR) 1 mode (combined) inj.: 5 µL sample: water extract of green tea gradient:

Min. % A %B LC-MS G003373 0.0 80 20 0.66 80 20 1.33 45 55 7 2.10 80 20 5 2 6 1. Gallocatechin 509th Injection 2. Caffeine 3. Epigallocatechin 3 4. Catechin 5. Epicatechin 6. Epigallocatechin gallate 7. Epicatechin gallate / 814-359-3041

1 (US and Canada only)

G003374

Complex Separation of Antiretrovirals on Ascentis RP-Amide with MS Detection The suitability of the short Ascentis RP-Amide column in the separation of drugs used in combination antiretroviral Ascentis RP-Amide therapy is demonstrated. These include nevirapine, a non- 3 nucleoside reverse transcriptase inhibitor; lamivudine, zidovudine, stavudine, nucleoside analog drugs; and the

HIV-protease inhibitors lopinavir and saquinavir. The 4 2 5 6 chromatographic conditions were developed from the USP 1 method for nevirapine.

column: Ascentis RP-Amide, 3 cm x 4.6 mm I.D., 3 µm particles (565317-U) mobile phase: gradient A = 0.1% ammonium acetate (pH 5.3 with acetic acid), B = acetonitrile ﬂ ow rate: 1.0 mL/min. (split to the mass spectrometer) temp.: 35 °C det.: MS, ESI (-) and (+) in full scan mode (combined) / 814-359-3441 technical service: 800-359-3041 inj.: 5 µL G003378 sample: as indicated in 100% 0.1% ammonium acetate (pH 5.3 with acetic acid) gradient: (US only) Min. %A %B 0.0 95 5 2.0 40 60 3.0 40 60 3.1 95 5

1. Lamivudine 2. Stavudine 3. Zidovudine 4. Nevirapine 5. Saquinavir 6. Lopinavir ordering: 800-247-6628 ordering:

Elevated HPLC Performance 8 Maximum Retentivity Better sensitivity and resolution

2 High surface area silica (450 m /gram) enhanced when analytes evaporate under higher organic and high phase density on the silica conditions. Utilizing high organic mobile phases promotes make Supelco’s Ascentis family of easier sample preparation due to solubility. Preparative columns highly retentive. chromatography procedures are improved by minimizing evaporation and reconstitution steps. The high retentivity High retentivity allows the chromatographer to run at advantage extends into highly aqueous mobile phases for higher organic solvent composition. This is ideal for LC-MS RP-Amide and C8, thus retaining even the most polar applications and can increase MS sensitivity. Ionization compounds beyond the void. efﬁ ciency and the accompanying MS signal are often Maximum Retentivity

TRADEMARKS: ACE - Advanced Chromatography Technologies; Ascentis, CHROMASOLV, Discovery, SUPELCOSIL, Suplex - Sigma-Aldrich; Genesis - Argonaut; BetaMax, Prism - Thermo Electron Corporation; Inertsil - GL Sciences; Luna, Prodigy, Synergi - Phenomenex; Nucleosil - Macherey-Nagel; Polaris - Varian, Inc; Symmetry, SymmetryShield, XTerra - Waters Corporation; Zorbax - Agilent Technologies sigma-aldrich.com/supelco

ASCENTIS™ HPLC COLUMNS 9 Orthogonal Selectivity Resolve difﬁ cult analytes

While good retention and high effi ciency can create larger differences in selectivity for resolving are important in separations, selectivity diffi cult pairs or confi rming identity. Nothing on the market is the most powerful parameter for rivals Ascentis C18 and RP-Amide in orthogonal selectivity. achieving separations. This orthogonal difference is illustrated clearly in the log k vs. log k plot. Typical column pairs show less scatter as Supelco had that in mind when designing the Ascentis compared to this plot. family. Move between the C18 and C8 when small differences in retention and selectivity are desired. The RP-Amide

Choices in Separations NOTE: Large scatter of Selectivity, often the most critical parameter in developing data points indicates greater selectivity separations, has been optimized for the Ascentis C18 / compared to C18. RP-Amide phase pair by means of our Surface-Optimized Technology. Orthogonal Selectivity The log kC18 versus log kRP-Amide plot illustrates the / 814-359-3041 powerful differences in selectivity (orthogonality) of the Ascentis C18 and Ascentis RP-Amide column pair. Furthermore, the orthogonality of the Ascentis C18 and Ascentis RP-Amide column pair is dramatically illustrated by the selectivity differences for nortriptyline relative to a (US and Canada only) hydrophobic marker (below).

Analysis of Tricyclic Antidepressants on Ascentis C18 and Ascentis RP-Amide

column: Ascentis C18,15 cm x 4.6 mm I.D., 5 µm particles (581324-U) 1. Uracil Ascentis RP-Amide, 15 cm x 4.6 mm I.D., 5 µm particles (565324-U) 2. Nortriptyline mobile phase: 35:65, 25 mM ammonium phosphate, pH 7 methanol 3. Toluene ﬂ ow rate: 1.2 mL/min. 4. Imipramine temp.: 35 °C 5. Amitriptyline det.: UV, 230 nm inj.: 10 µL 2 2 NOTE: Elution order Ascentis C18 Ascentis RP-Amide is reversed for peaks 2 and 3. 3

1 1 3 / 814-359-3441 technical service: 800-359-3041

4 (US only)

4 5

G003381 G003382 ordering: 800-247-6628 ordering:

Elevated HPLC Performance 10 Applications

Steroids Ascentis RP-Amide shows strong afﬁ nity for 1. Estriol phenolic molecules, hence the increased 2. β-estriol α retention time for all four compounds. In 3. -estriol 4. Estrone this case, the C18 yields better selectivity and a faster run time due to dominance of hydrophobic interactions. Both columns ex- Ascentis C18 Ascentis RP-Amide hibit excellent peak shape for steroids. Many samples can be analyzed equally well with Ascentis C18, C8, and RP-Amide.

column: Ascentis C18, 15 cm x 4.6 mm I.D., 5 µm particles (581324-U) Ascentis RP-Amide, 15 cm x 4.6 mm I.D., 5 µm particles (565324-U)

Applications mobile phase: 55:45, water:acetonitrile ﬂ ow rate: 1 mL/min. temp.: 35 °C det.: UV at 220 nm inj.: 10 µL G002330 G002672

Pharmaceutical Bases The separation of acetaminophen and 1. Acetaminophen pseudoephedrine illustrates the dramatic 1 2. Pseudoephedrine 2 1 effect of simply changing the stationary phase. The ability of Ascentis RP-Amide to shield pseudoephedrine, a strong base, from silanols causing less retention and reversal Ascentis C18 Ascentis RP-Amide of elution order is an important property of the embedded polar group phase. 2 column: Ascentis C18, 15 cm x 4.6 mm I.D., 5 µm particles (581324-U) Ascentis RP-Amide, 15 cm x 4.6 mm I.D., 5 µm particles (565324-U) mobile phase: 15:85, acetonitrile: 25 mM potassium phosphate ﬂ ow rate: 1 mL/min. temp.: 35 °C det.: UV at 214 nm 024 inj.: 10 µL G003363Min G003364

Tricyclic Antidepressants (TCAs) The TCAs with a secondary amine show en- 1. Nordoxepin hanced retention on Ascentis RP-Amide due 2. Protriptyline to hydrogen bonding interactions, whereas 3. Nortriptyline 4. Impramine the methylated amines show higher retention 5. Amitriptyline on Ascentis C18 because they are more 1 hydrophobic. Ascentis C18 Ascentis RP-Amide 1 3 column: Ascentis C18, 3 4 15 cm x 4.6 mm I.D., 5 µm particles (581324-U) Ascentis RP-Amide, 15 cm x 4.6 mm I.D., 5 5 µm particles (565324-U) 2 mobile phase: 30:70, 25 mM ammonium 2 4 phosphate, pH 7: methanol 5 ﬂ ow rate: 1 mL/min. temp.: 35 °C det.: UV at 254 nm inj.: 10 µL

sigma-aldrich.com/supelco G003371 G003372

ASCENTIS™ HPLC COLUMNS 11

Polar Molecules Under Highly Aqueous Conditions

Ascentis C8 often yields enhanced retention 1. L-tartaric 2. Lactic Acid than Ascentis C18 for small polar molecules 1 1 under highly aqueous conditions. Greater Ascentis C183. Citric Acid Ascentis C8 4. Fumaric Acid retention for Ascentis C8 may be related to 5. Acrylic Acid greater wettability of Ascentis C8 as com- 2 4 pared to Ascentis C18. 3 4 2 3 5 column: Ascentis C18, 15 cm x 4.6 mm I.D., 5 µm particles (581324-U) Ascentis C8, 15 cm x 4.6 mm I.D., 5 µm particles (581424-U) 5 mobile phase: 3:97, methanol:water with 0.1% TFA ﬂ ow rate: 1 mL/min.

temp.: 35 °C Applications det.: UV at 220 nm inj.: 10 µL

G003365 G003368

Metabolite Studies / 814-359-3041 Small polar molecules are often important 1. L-tartaric 2. Lactic Acid in degradant and metabolite studies. This 1 Ascentis C81 Ascentis RP-Amide example illustrates the suitability of both 3. Citric Acid 4. Fumaric Acid Ascentis RP-Amide and C8 for small polar 4 5. Acrylic Acid 2 5 molecules. Furthermore, a selectivity change 4 is realized with fumaric acid due to increased (US and Canada only) 2 3 3 ability to hydrogen bond with the amide 5 group.

column: Ascentis C8, 15 cm x 4.6 mm I.D., 5 µm particles (581424-U) Ascentis RP-Amide, 15 cm x 4.6 mm I.D., 5 µm particles (581324-U) mobile phase: 3:97, methanol:water with 0.1% TFA ﬂ ow rate: 1 mL/min. temp.: 35 °C det.: UV at 220 nm inj.: 10 µL G003368 G003367

Pharmaceutical Bases at pH 7

Analysis of bases at neutral pH often yields 1. Desmethyl Doxepin enhanced retention over acidic mobile phase 2. Protriptyline but sometimes causes poor peak shape 3. Desimpramine 1 1 3 with C18 and C8 columns due to silanol 4. Nortriptyline 3 interaction. In this example, a mix of tricyclic 5. Doxepin 6. Impramine antidepressants at pH 7 shows excellent peak 7. Amitriptyline 4 shape on both Ascentis C8 and C18. Under 4 8. Trimpramine / 814-359-3441 technical service: 800-359-3041 these conditions, the increased hydrophobic retentivity of Ascentis C18 versus C8 is appar- ent, especially in the late eluting compounds. Ascentis C18 Ascentis C8 (US only) 2 2 5 5 6 column: Ascentis C18, 6 7 15 cm x 4.6 mm I.D., 7 5 µm particles (565324-U) 8 8 Ascentis C8, 15 cm x 4.6 mm I.D., 5 µm particles (581424-U) mobile phase: 30:70; 25 mM ammonium phosphate, pH 7.0: methanol ﬂ ow rate: 1.5 mL/min. temp.: 35 °C det.: UV at 254 nm G003369 G003370 800-247-6628 ordering: inj.: 20 µL

Elevated HPLC Performance 12 Column Selection Guide

Supelco offers one of the most diverse sets of HPLC phases The Discovery line introduced the versatile Discovery HS F5 available from any company. The following guide displays and the Zr series for high temperature and extreme pH Ascentis and spans our 25-year history of excellence in HPLC conditions as well as the BIO wide series for proteins and columns. SUPELCOSIL LC-18-DB was the industry’s ﬁ rst base- peptides. deactivated column and is still popular in HPLC methods.

Primary USP Particle Pore Surface pH Carbon Column Feature Beneﬁ ts Applications Code Size Size Area Range Load Ascentis High Retentivity Strong retention, General reversed L1 3,5,10 µm 100 Å 450 m2/g 2-8 25% C18 Increased retentivity, phase; Hydrophobic High carbon loading & polar compounds; & proprietary Small molecule endcapping preparative Discovery Intermediate Analyze a wide Scouting wide L1 5 µm 180 Å 200 m2/g 2-8 12% C18 Pore Size range of molecular range in molecular weights (small weights; General molecules to use C18 biomolecules) SUPELCOSIL First Deactivated Long history Basic compounds, L1 3,5 µm 120 Å 300 m2/g 2-8 11% LC-18-DB LC Column of performance Less retentive than & reproducibility Ascentis C18; General use C18 Ascentis Excellent Strong retention; General reversed L7 3,5,10 µm 100 Å 450 m2/g 2-8 15%

Column Selection Guide C8 Peak Shape Increased retentivity; phase; Hydrophobic high carbon loading & polar compounds; & proprietary Small molecule endcapping preparative Discovery Wide Pore Separates peptides, Peptides, proteins various 3,5,10 µm 300 Å 100 m2/g 2-8 various% BIO Wide Size proteins, & other proteomics & Series large molecules proteolytic digests; (>2000 MW); Scalable from Variety of hydro- analytical to phobicities preparative Ascentis Alternate Perfect choice for Excellent selectivity L60 3,5,10 µm 100 Å 450 m2/g 2-8 19% RP-Amide Selectivity method development for acids, bases, and peak identity & polar compounds, confi rmation; especially phenols Low bleed ideal for and amines; LC-MS applications Small molecule preparative Discovery High Polarity Suitable for Polar compound L10 5 µm 180 Å 200 m2/g 4-8 4.5% CN reversed phase as separations well as normal including phase; Unique carboxylic acids selectivity for & amines polar & aromatic compounds Discovery Versatile retention Excellent for basic Hydrophobic & L43 3,5,10 µm 120 Å 300 m2/g 2-8 12% HS F5 mechanism, drug separations, ion exchange Pentafl uorophenyl Effi cient separation mechanisms create phase of bases at high unique selectivity organic mobile phase for basic (~90% ACN) molecules Discovery Zirconia; Stable for acids pH range of 1-13; various 3,5 µm 300 Å 30 m2/g 1-13 various% Zr Wide pH range/ & bases, allows Temp. range up to Series high temperature for use of full pH 100 °C (200 °C w/ spectrum; special hardware); High temperature Alternate selectivity capabilities from ion exch. mech. SUPELCOSIL Ion Ion exchange Aminopropyl phase L9 & 5 µm 120 Å 170 m2/g 2-8 n/a SCX Separations columns for separation of L52/ SAX anions; L14 Propylsulfonic acid groups for separating cations sigma-aldrich.com/supelco

ASCENTIS™ HPLC COLUMNS 13 and Accessories / 814-359-3041 Ascentis Tools, Reagents Ascentis Tools, (US and Canada only)

Reagents and Accessories

Acids, Bases, and Buffers spectrophotometry, environmental testing, and other The quality and reproducibility of liquid chromatography analytical applications. CHROMASOLV Plus solvents separations depend not only on instrumentation and undergo rigorous specifi cation testing to provide you columns but also on the quality of the reagents that are with lot-to-lot consistency. used. The acids, bases, and buffers offered by Supelco have been optimized for liquid chromatography applications. HPLC Autosampler Vials Each is lot tested and precisely controlled to meet the high Supelco offers the most popular vial styles for LC auto- standards necessary for successful HPLC separations. samplers in both clear and amber glass. A variety of seals or caps is available for all vials. ABC vials with a 9 mm fi nish CHROMASOLV® LC-MS Grade Solvents can be used for applications requiring a tight seal where you These LC-MS solvents are designed specifi cally with low do not want to use a hand crimper. This vial has the thread / 814-359-3441 technical service: 800-359-3041 content (≤100 ppb) of alkaline impurities such as calcium, higher on the glass to allow it to be used with robotic magnesium, potassium, and sodium, which can interfere in autosamplers. The 6.0 mm large opening fi nish has made (US only) your analysis by forming adducts with the analyte. The this vial very popular. Crimp top vials are recommended for CHROMASOLV LC-MS solvents are also run through specifi c samples where a tight seal is required. These are available UV-spectroscopic quality control tests to guarantee the with either a standard (4.6 mm) or large (6.0 mm) opening. traditional CHROMASOLV specifi cations. Snap top vials can be used with non-volatile samples where the seal isn’t critical. Many customers prefer this style to CHROMASOLV Plus HPLC Grade Solvents avoid using a crimper and decapper. Screw top vials are also These are specifi cally designed to be your multipurpose, available in both a standard or large opening style. Screw top styles provide a good seal and are easy to use.

high-purity solvents. They are tested for suitability in HPLC, 800-247-6628 ordering:

Elevated HPLC Performance 14

“We are committed to the success of our Customers, Employees, and Shareholders through leadership in Life Science, High Technology, and Service.” The Service You Deserve… Easy Ordering On-Time Delivery Supelco offers many ways to easily place your order or Still waiting for HPLC or capillary GC columns that you inquire about an existing order. You can call, fax, or visit our ordered last week or last month? Not getting responsive web site 24 hours a day, 7 days a week. customer service? Get the service you deserve today with In the USA and Canada, please contact us at: products that meet your performance demands and are Phone : 800-247-6628 / 800-325-3010 / delivered when you need them. 814-359-3441 Fax: 800-325-5052 / 814-359-5459 Superior Support Web: sigma-aldrich.com/supelco When you want additional information about a product, Other countries, contact your local Sigma-Aldrich or help with a problem, our knowledgeable Technical representative. For more information, visit sigma-aldrich.com Service chemists and our extensive library of free technical

Customer Service Ready to order products? Searching for Material Safety literature are just a telephone call, fax message, or email Data Sheets or other product information? Need a new away. We are always ready to respond to your questions, catalog? Many of your questions can be answered at our suggestions, criticisms, and changing needs. web site. Go to sigma-aldrich.com/supelco to search for In the USA and Canada, please contact us at the products, check up-to-the-minute availability, place orders, following telephone numbers Monday - Friday during the check order status, or directly trace your order with a hours of 8AM - 8PM Eastern Time tracking number that is hot linked to the carrier’s web site. Phone: 800-359-3041 / 814-359-3041 You can check the status and trace an order in two clicks! Fax: 800-359-3044 / 814-359-5468 Email: [email protected] Other countries, contact your local Sigma-Aldrich representative. For more information, visit sigma-aldrich.com …from the Company You Trust sigma-aldrich.com/supelco

ASCENTIS™ HPLC COLUMNS 15

Ordering Information

ID (mm) Length (cm) Particle Size (µm) C18 RP-Amide C8 Ascentis HPLC Columns 1.0 5 3 581311-U 565309-U 581412-U 1.0 10 3 581364-U 565389-U 581435-U 1.0 15 3 581365-U 65566-U 581436-U 2.1 2 3 581312-U 565313-U 581413-U 2.1 2 5 581368-U 565391-U 581439-U 2.1 3 3 581313-U 565314-U 581414-U 2.1 3 5 581327-U 565331-U 581430-U 2.1 5 3 581300-U 565300-U 581400-U 2.1 5 5 581303-U 565303-U 581420-U 2.1 10 3 581301-U 565301-U 581401-U 2.1 10 5 581326-U 565304-U 581419-U 2.1 15 3 581302-U 565302-U 581402-U 2.1 15 5 581304-U 565305-U 581421-U 2.1 25 5 581305-U 565306-U 581422-U 3.0 2 3 581314-U 565315-U 581415-U 3.0 2 5 581328-U 565332-U 581431-U 3.0 3 3 581302-U 565310-U 581403-U 3.0 3 5 581369-U 565392-U 581440-U 3.0 5 3 581307-U 565311-U 581404-U 3.0 5 5 581329-U 565333-U 581432-U 3.0 10 3 581308-U 565312-U 581405-U 4.6 2 3 581315-U 565316-U 581416-U 4.6 2 5 581330-U 565335-U 581433-U 4.6 3 3 581316-U 565317-U 581417-U 4.6 3 5 581331-U 565336-U 581434-U 4.6 5 3 581320-U 565320-U 581406-U 4.6 5 5 581323-U 565323-U 581423-U Information Ordering 4.6 10 3 581321-U 565321-U 581407-U / 814-359-3041 4.6 10 5 –––––––– 565328-U –––––––– 4.6 15 3 581322-U 565322-U 581408-U 4.6 15 5 581324-U 565324-U 581424-U 4.6 15 10 581350-U 565352-U –––––––– 4.6 25 5 581325-U 565325-U 581425-U 4.6 25 10 581351-U 565353-U –––––––– 10.0 1 5 –––––––– 565376-U –––––––– 10.0 1 10 581388-U –––––––– –––––––– (US and Canada only) 10.0 5 5 581340-U 565340-U –––––––– 10.0 5 10 581352-U 565354-U –––––––– 10.0 10 5 581341-U 565341-U –––––––– 10.0 10 10 581353-U 565355-U –––––––– 10.0 15 5 581342-U 565343-U –––––––– 10.0 15 10 581354-U 565356-U –––––––– 10.0 25 5 581343-U 565344-U –––––––– 10.0 25 10 581355-U 565357-U –––––––– 21.2 5 5 581344-U 565345-U –––––––– 21.2 5 10 581356-U 565358-U –––––––– 21.2 10 10 581357-U 565359-U –––––––– 21.2 15 10 581358-U 565360-U –––––––– 21.2 25 5 581347-U 565348-U –––––––– 21.2 25 10 581359-U 565361-U ––––––––

Ascentis Supelguard Cartridges Kit 2.1 2 3 581376-U –––––––– –––––––– Pack of 2 2.1 2 3 581377-U –––––––– –––––––– Pack of 2 2.1 2 5 581370-U 565372-U –––––––– Kit 2.1 2 5 581371-U 565373-U –––––––– Pack of 2 3.0 2 5 581374-U 565374-U –––––––– Kit 3.0 2 5 581375-U 565375-U –––––––– Kit 4.0 2 3 581378-U –––––––– –––––––– Pack of 2 4.0 2 3 581379-U –––––––– –––––––– Pack of 2 4.0 2 5 581372-U 581370-U 581426-U Kit 4.0 2 5 581373-U 581371-U 581427-U

Kits include one cartridge, stand alone holder, a piece of tubing, 2 nuts and 2 ferrules. / 814-359-3441 technical service: 800-359-3041

Ascentis Validation Packs (US only) 4.6 15 5 581390-U 565394-U –––––––– 4.6 25 5 581391-U 565395-U –––––––– Validation Packs include 3 columns, each from a different bond lot.

Custom Columns If you don’t see the column dimension you need, we can help. Contact our Technical Service group at 800-359-3041 / 814-359-3041 or by email at [email protected]. ordering: 800-247-6628 ordering:

Elevated HPLC Performance Argentina France Japan Singapore SIGMA-ALDRICH DE ARGENTINA, Sigma-Aldrich Chimie S.à.r.l. SIGMA-ALDRICH JAPAN K.K. Sigma-ALDRICH PTE. LTD. S.A. Tel appel gratuit: 0800 211 408 Tokyo Tel: 03 5796 7300 Tel: 65-67791200 Tel: 54 11 4556 1472 Fax appel gratuit: 0800 031 052 Tokyo Fax: 03 5796 7315 Fax: 65-67791822 Fax: 54 11 4552 1698 Germany Korea South Africa Australia Sigma-Aldrich Chemie GmbH SIGMA-ALDRICH KOREA Sigma-Aldrich Sigma-Aldrich Pty., limited Free Tel: 0800-51 55 000 Tel: 031-329-9000 South Africa (Pty) Ltd. Free Tel: 1800 800 097 Free Fax: 0800-649 00 00 Fax: 031-329-9090 Free Tel: 0800 1100 75 Free Fax: 1800 800 096 Free Fax: 0800 1100 79 Tel: 612 9841 0555 Greece Malaysia Tel: 27 11 979 1188 Fax: 612 9841 0500 SIGMA-ALDRICH (o.m.) Ltd SIGMA-ALDRICH (M) Sdn. Bhd Fax: 27 11 979 1119 Tel: 30 210 9948010 Tel: 603-56353321 Austria Fax: 30 210 9943831 Fax: 603-56354116 Spain SIGMA-ALDRICH Handels GmbH SIGMA-ALDRICH QUÍMICA S.A. Tel: 43 1 605 81 10 Hungary Mexico Free Tel: 900 101376 Fax: 43 1 605 81 20 SIGMA-ALDRICH Kft SIGMA-ALDRICH QUÍMICA, S.A. de C.V. Free Fax: 900 102028 Tel: 06-1-235-9054 Free Tel: 01-800-007-5300 Tel: 91 661 99 77 Belgium Fax: 06-1-269-6470 Free Fax: 01-800-712-9920 Fax: 91 661 96 42 Sigma-Aldrich NV/SA. Ingyenes zöld telefon: 06-80-355-355 Free Tel: 0800-14747 Ingyenes zöld fax: 06-80-344-344 The Netherlands Sweden Free Fax: 0800-14745 Sigma-Aldrich Chemie BV SIGMA-ALDRICH SWEDEN AB Tel: 03 899 13 01 India Tel Gratis: 0800-0229088 Tel: 020-350510 Fax: 03 899 13 11 SIGMA-ALDRICH CHEMICALS Fax Gratis: 0800-0229089 Fax: 020-352522 PRIVATE LIMITED Tel: 078-6205411 Outside Sweden Tel: +46 8 7424200 Brazil Telephone Fax: 078-6205421 Outside Sweden Fax: +46 8 7424243 Sigma-Aldrich Brasil Ltda. Bangalore: 91-80-5112-7272 Tel: 55 11 3732-3100 New Delhi: 91-11-5165 4255 New Zealand Switzerland Fax: 55 11 3733-5151 Mumbai: 91-22-2570 2364 SIGMA-ALDRICH PTY., LIMITED SIGMA-ALDRICH CHEMIE LLC Hyderabad: 91-40-5584 5488 Free Tel: 0800 936 666 Swiss Free Call: 0800 80 00 80 Canada Fax Free Fax: 0800 937 777 Tel: +41 81 755 2828 Sigma-Aldrich Canada Ltd. Bangalore: 91-80-5112-7473 Tel: 61 2 9841 0500 Fax: +41 81 755 2815 Free Tel: 800-565-1400 New Delhi: 91-11-5165 4266 Fax: 61 2 9841 0500 Free Fax: 800-265-3858 Mumbai: 91-22-2579 7589 United Kingdom Tel: 905-829-9500 Hyderabad: 91-40-5584 5466 Norway Sigma-Aldrich Company Ltd. Fax: 905-829-9292 SIGMA-ALDRICH NORWAY AS Free Tel: 0800 717181 Ireland Tel: 23 17 60 60 Free Fax: 0800 378785 China Sigma-Aldrich Ireland Ltd. Fax: 23 17 60 50 Tel: 01747 833000 SIGMA-ALDRICH CHINA INC. Free Tel: 1800 200 888 Fax: 01747 833313 Tel: 86-21-6386 2766 Free Fax: 1800 600 222 Poland SAFC (UK): 01202 712305 Fax: 86-21-6386 3966 Tel: 353 1 4041900 SIGMA-ALDRICH Sp. z o.o. Fax: 353 1 4041910 Tel: 061 829 01 00 United States Czech Republic Fax: 061 829 01 20 SIGMA-ALDRICH SIGMA-ALDRICH s.r.o. Israel P.O. Box 14508 Tel: +420 246 003 200 Sigma-Aldrich Israel Ltd. Portugal St. Louis, Missouri 63178 Fax: +420 246 003 291 Free Tel: 1-800-70-2222 Sigma-Aldrich QuÍmica, S.A. Toll-free: 800-325-3010 Tel: 08-948-4100 Free Tel: 800 202180 Call Collect: 314-771-5750 Denmark Fax: 08-948-4200 Free Fax: 800 202178 Toll-Free Fax: 800-325-5052 Sigma-Aldrich Denmark A/S Tel: 21 9242555 Tel: 314-771-5765 Tel: 43 56 59 10 Italy Fax: 21 9242610 Fax: 314-771-5757 Fax: 43 56 59 05 SIGMA-ALDRICH S.r.l. Internet: Telefono: 02 33417310 Russia sigma-aldrich.com Finland Fax: 02 38010737 SIGMA-ALDRICH RUS, LLC Sigma-Aldrich Finland Numero Verde: 800-827018 Tel: +7 (095) 975-1917/3321 Tel: (09) 350 9250 Fax: +7 (095) 975-4792 Fax: (09) 350 92555

Order/Customer Service 800-247-6628, 800-325-3010 Technical Service 800-359-3041, 814-359-3041 Fax 800-325-5052 ● E-mail [email protected] Fax 800-359-3044, 814-359-5468 ● E-mail [email protected] SUPELCO ● 595 North Harrison Road, Bellefonte, PA 16823-0048 ● 814-359-3441

We are committed to the success of our Customers, Employees and Shareholders through leadership in Life Science, High Technology and Service. ® ™ ® ® The SIGMA-ALDRICH Family sigma-aldrich.com/supelco ©2005 Sigma-Aldrich Co. Printed in USA. Supelco brand products are sold through Sigma-Aldrich, Inc. Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see reverse side of the invoice or packing slip. Supelco is a registered trademark of Sigma-Aldrich Co. HLV T404114B