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Ordering Information OmniSpher C - ChromSep Stainless Steel cartridge columns Dimensions L * ID Particle size Complete Replacement Replacements Guardcolumns (mm) (µm) System Cat.no. Cat.no Cat.

1 Ordering Information OmniSpher C - ChromSep Stainless Steel cartridge columns Dimensions L * ID Particle size Complete Replacement Replacements Guardcolumns (mm) (µm) System Cat.no. Cat.no Cat.no (set of ) Cat.no (set of ) 0 *.0 CP CP CP 0 *.0 CP CP CP 0 *.0 CP CP CP 00 *.0 CP9 CP0 CP CP 0 *.0 CP CP CP9 0 *.0 CP0 CP CP CP 00 *.0 CP CP CP CP 0 *. CP CP CP CP 00 *. CP0 CP CP CP 00 *.0 CP0 CP CP CP 0 *.0 CP0 CP CP CP 0 *.0 CP09 CP CP9 CP 00 *. CP0 CP0 CP CP 0 *. CP0 CP CP CP 0 *. CP0 CP9 CP CP Complete system includes: ChromSep hardware, OmniSpher columns and guard column. Everything to start your analysis. OmniSpher C - Conventional Valco columns Dimensions L * ID Particle size Column Chromguard Chromguard (mm) (µm) Cat.no. Complete Replacements (set of ) 00 *.0 CP0 CP090 CP90 00 *. CP CP090 CP90 0 *.0 CP CP090 CP90 0 *.0 CP CP090 CP90 00 *. CP CP0 CP 0 *. CP CP0 CP 0 *. CP CP0 CP Varian Analytical Instruments, serving worldwide markets in: Agriculture Basic Chemical Biotechnology Clinical Electronics Environmental Photonics Toxicology Pharmaceutical Food and Beverage Metals and Mining Petroleum and Petrochemical Varian Sales Offices Varian is committed to a process of continuous improvement which demands that we understand and then meet or exceed the needs and expectations of our customers both inside and outside the company in everything we do. Varian OmniSpher C columns UNIVERSAL OmniSpher R P L C C O L U M N Varian Analytical Instruments North America: , Europe The Netherlands: Asia Pacific Australia:.90. Latin America Brazil:...0 Other sales offices and dealers throughout the world GC GC/MS AAS ICP ICP-MS UV-Vis-NIR Fluorescence NMR Analytical Supplies MDL000R0LU000M

OmniSpher The universal RPLC column Speed up your analysis with OmniSpher C ChromSep cartridge system for analytical columns Varian OmniSpher C columns, using an improved universal stationary phase

widely used column packings in are silica based stationary phases derivatized with octadecyl (C, ODS) groups. These packings, however, have their limitations.

Stationary phase The OmniSpher C is a newly developed column for reversed phase analysis and is based on very pure silicagel with controlled and extremely low concentrations of metals and salts.

2 OmniSpher The universal RPLC column Speed up your analysis with OmniSpher C ChromSep cartridge system for analytical columns Varian OmniSpher C columns, using an improved universal stationary phase for reverse phase, offer the following advantages: One column does the job Faster method development Turns your research lab into a routine lab Superior performance, inertness and stability The most widely used column packings in are silica based stationary phases derivatized with octadecyl (C, ODS) groups. These packings, however, have their limitations. For instance, their limited ph range causes difficulties in analyzing polar, acidic and basic compounds, and their adsorption and irreproducibility cause troublesome peak-tailing. Stationary phase The OmniSpher C is a newly developed column for reversed phase analysis and is based on very pure silicagel with controlled and extremely low concentrations of metals and salts. The spherical particles allow the packing of highly-efficient columns having excellent permeability and good mechanical strength without forming fines during the packing procedure. Varian has developed a single step derivatization procedure for its Chrompack columns. This procedure allows chemical bonding of monofunctional C groups to the silica surface. It is based on a high purity silicagel with controlled minimal impurity concentration of metals and salts. OmniSpher can be used for neutral, polar, acidic and basic compounds. That makes OmniSpher a true universal reverse phase packing. The applications shown in this brochure will convince you. One column does do the job Today s routine Quality Control laboratories require accurate analysis of more samples with fewer resources. Methods must be easy to set up, simple to operate and require a minimum of maintenance. You can work flexibly and improve productivity with the uniquely inert OmniSpher C columns because they are applicable for almost any component. Your chromatography results will benefit from improved quantification through increased selectivity. Column Quality Assurance test Every OmniSpher column is individually tested with a specially developed test mixture to guarantee the specifications of each column before shipment. Standard test parameters are: the plate number Nth/ (reduced) plate height h, peak asymmetry factor As, retention factor k, separation factor a and the column pressure drop Dp and flow resistance factor F. Typical specifications for a cm x. mm ID standard OmniSpher C column are as follows: Chromatographic characteristics of. Partical size µm Retention factor k o-terphenyl 9. Separation factor /. Theoretical plate number N th >0,000/m Plate height H < µm Reduced plate height h <. Asymmetry factor 0.<As<. Flow resistance factor F Why choose for OmniSpher columns? Unsurpassed column performance High batch-to-batch reproducibility Excellent stability and reproducibility of separation and retention times. High-efficiency and high-speed screening Ideal for LC-MS with a broad range of short.0mm ID columns Excellent column stability Long column liftime due to stable bonding technology High separation power High retention-time stability up to 0,000 column volumes (ph.0) Universal column inertness Ideal for neutral, polar, acidic and basic compounds Columns are eluted with very symmetrical peaks. Exellent choice for the analysis of basic drugs Each column is individually validated with a new, specific test method No tailing peaks Excellent symmetrical peaks Significantly better retention time and repeatability. It is well know that by reducing the particle size of the column packing, the column efficiency and the separation power of the column (resolution) increase. But this has to be paid for with a substantial increase of the column pressure drop (almost times higher). In practice it means that most routine applications will be performed with shorter µm columns, typically between and 0 cm column length. An important theoretical factor is that the higher performance occurs with shorter analysis times (higher flow velocities/ analysis speed) and smaller particle size. The maximum resolution shifts to higher speeds. In practice the flow rate can be increased to take full advantage of smaller particles packed in a shorter column. These relations can be derived from the plate height/velocity relation in the H-u curve as shown in the figure. Changing from to micron particle columns with OmniSpher C results in more efficient separation and/or higher speed (depending on the column length of choice), but with exactly the same selectivity. It allows you to choose or switch to the particle size most suited to your application. For high-resolution separation, a longer column (0 to cm) with micron particles can be applied. For fast separation of a limited number of components to be quantified (typically - peaks), short columns ( - 0 cm) can be used with an analysis time of - 0 minutes. - 0 mm conventional columns for robust chromatography - 0 and 00 mm columns for the optimum ratio between speed and efficiency - 0, 0 and 0 mm ultra-fast columns for screening techniques, e.g. LC-MS and LC-MS-MS High throughput screening / fast LC-MS analysis In pharmaceutical drug development and high throughput screening, often only one peak has to be separated from the sample matrix. In LC-MS applications, also the resolution between the peaks of interest is less important then with classical UV-detection. In all these cases it is possible to operate with very short columns of, or cm length. Furthermore, if these columns are designed with a mm ID flow rates between 0. and 0. ml/min are possible. This does not only decrease solvent consumption and sample dilution in the column, but is also exactly in the optimum flow range of many modern LC-MS systems. Varian's short columns packed with OmniSpher C are the columns of choice for your fast separations, especially for screening and analyzing basic drugs and other polar compounds. -.0 and. mm ID conventional columns for robust chromatography -.0 mm solvent saving columns -.0 mm ID columns for solvent saving, increased sensitivity and LC-MS Typical specifications for a 0 cm x. mm ID OmniSpher C column are: Chromatographic characteristics of OmniSpher C. Partical size µm Retention factor k o-terphenyl 9. Separation factor /. Theoretical plate number N th >0,000/m Plate height H <.µm Reduced plate height h <. Asymmetry factor 0.<As<. Flow resistance factor F The reliable, easy-to-use ChomSep cartridge system is now available with most phases, including the new generation reversed phase OmniSpher C columns from Varian. The system combines simplicity with extraordinary flexibility and considerable savings on column and operation costs. The flexibility of the ChromSep column system The ChromSep system allows you to select the right column for your application. If you want a robust column for standard separations select a 0 x. mm column, but for very fast separation or for screening techniques, chose a 0, 0 or 0 mm long column. And when selecting the column ID you can choose a conventional robust. or.0 mm column or, if you prefer to save on solvents and increase sensitivity, select a.0 or.0 mm ID. These.0 mm ID columns, by the way, are highly suited for LC-MS applications and screening techniques. Durable column hardware for cost savings and easy column replacement Since ChromSep column housings are available in lengths of 0 0, 0, 00, 0, 00, 0 and 00 mm, you can use any length or combination of cartridge columns. Match the column length with the separation you need and minimize your analysis time. The ChromSep durable SS column holder is a one time investment. You can easily replace the cartridge column without the need for tools. Unlike conventional stainless steel columns, you don t have to throw away the column hardware and that s good for your budget and the environment. Integrated protection for long column life The ChromSep column s ingenious design allows you to use a guard column without introducing any dead volume. By directly fitting the guard column onto the top of the analytical column in the same holder, the analytical performance of the column is fully guaranteed. In addition, a removable screen on top of the frit protects your analytical column. This screen can easily be replaced (if it becomes contaminated) without damage to the packing because the frit remains in place. We guarantee validated performance for the best results Every step in the manufacture and testing of ChromSep columns is critically controlled and monitored. Each column is characterized with specifications for key chromatographic flexibility parameters. fits your We guarantee application that you only use columns which perform to the tightest and highest operation specifications. We ensure that methods can be reproduced, column after column, batch after batch and from laboratory-to-laboratory. The ChromSep column: The ChromSep column: flexibility fits your application End head and capillary connector enable fast and easy column replacement without tools and without disconnecting tubing. Glass as well as stainless steel replacement columns available Engraved tracking information with column description and unique serial number for validation protocols Various selective packings to optimize your application Highly efficient packed replacement columns, individually tested, available in economic multi-packs Screen, easily replaceable without damaging the packing, removes dirt for longer column lifetime Smaller is Faster Integrated guard column for zero dead volume connection without efficiency loss, available in economic -packs

3 Technical information Techn ical information Technical i formation Technical inform tion Technical information echnical information Techni cal information Technical in formation Technical inform tion Technical information echnical information Techni cal information Technical in formation Technical inform tion Technical information OmniSpher C columns Technical information Techn ical information Technical i UNIVERSAL OmniSpher R P L C C O L U M N

OmniSpher : Hi-tech universal stationary phase Superior column performance The mechanical and chromatographic properties of OmniSpher C have allowed Varian to develop breakthrough RPLC columns that

4 OmniSpher : Hi-tech universal stationary phase Superior column performance The mechanical and chromatographic properties of OmniSpher C have allowed Varian to develop breakthrough RPLC columns that offer a high degree of reproducibility. Typical plate counts for a cm x. mm ID column are 0,000-90,000 plates/meter. Column inertness Several tests are regularly performed to measure the inertness and universal applicability of the new OmniSpher columns. Basic and polar compounds are always selected for column tests to study the inertness of the stationary phase: Mixture of phenol and caffeine To check the hydrogen bonding capacity of the residual silanol groups Peak symmetry of caffeine and it s relative retention to phenol is an indication of silanol activity Mixture of phenol and benzylamine to check the ion exchange capacity and dissociation of free silanols when changing from low to high ph. OmniSpher C shows substantially smaller peak shifts than a comparable commercial reversed phase packing material. Procainamide and N-Acetylprocainamide (pka values above 9) At ph. the symmetry factors for both compounds are almost ideal. Varian's OmniSpher C columns deliver excellent results when compared to highly qualified phases. Tricyclic antidepressants The chromatogram indicates that the symmetry factor As of amitriptyline (USP tailing factor) is only., which is excellent under these mild mobile phase conditions. This superior performance explains why the OmniSpher C phase is a good choice for analyzing basic drugs. Basic test mixture According to McCalley, a text mixture containing a range of amines and other basic compounds is tested under standard conditions. At ph, the peak shape of all components is excellent. Under these conditions, some peaks show tailing on columns with regular RPLC packings due to protonation of the stationary phase. With OmniSpher C this effect is not visible. Tricyclic Antidepressants on. Uracil. Protriptyline. Nortriptyline. Doxepin. Imipramine. Amitriptyline USP tailing factor =.

Column stability The chromatographic stability of OmniSpher C is regularly tested by continuously pumping freshly prepared mobile phases of methanol/0 mmol phosphate buffer = 0 / 0 (v/v) at three

A mixture with tricyclic antidepressants is used to test the plate count, retention factor, selectivity factor and peak symmetry after every,000 column volumes of pumped eluent.

5 Column stability The chromatographic stability of OmniSpher C is regularly tested by continuously pumping freshly prepared mobile phases of methanol/0 mmol phosphate buffer = 0 / 0 (v/v) at three different ph values (.0,. and.0). A mixture with tricyclic antidepressants is used to test the plate count, retention factor, selectivity factor and peak symmetry after every,000 column volumes of pumped eluent. This is continued for 0,000 column volumes. Column efficiency A typical van Deemter plot of a packed column (0 x. mm ID) shows a very shallow slope at the right part of the curve, even at flow rates - times above the optimum. According to Prof. Th. Welsch (University of Ulm, Germany), this indicates excellent mass-transfer characteristics of the stationary phase. Therefore high-speed analysis, such as in LC-MS, is possible without jeopardizing column efficiency. The reduced plate height h in the van Deemter optimum is below, which is excellent. Hence the efficiency of the column is very high. pressure drop p [bar] reduced plate height h The data and graphs from the neutral and acidic endurance test show virtually no significant changes in performance. The basic endurance test shows some small changes in performance of the efficiency and peak symmetry. Retention factors and selectivity coefficients in this test do not change significantly, indicating a good chemical stability. Under standard conditions, the OmniSpher C's stability toward the ph of the mobile phase is specified in the range of.0 < ph < mobile phase velocity [mm/s] mobile phase flow rate [mm/s]

6 Applications A wide range of compounds with different polarities and pka values has been used to determine the universal applicability of OmniSpher C columns. These examples include neutral, polar, acidic and basic compounds in different application fields. UNIVERSAL OmniSpher R P L C C O L U M N Pharmaceuticals and drugs Anti epileptics OmniSpher C successfully separated anti-epileptic drugs using an acetonitrile/water gradient without buffer or other additives in the mobile phase. Consistency of the analysis can be improved by adding a mild buffer (0.0 M) to the mobile phase using a 0 cm column with isocratic elution. Antidepressants An excellent peak shape for these basic drugs is obtained at a ph of.. Due to the high selectivity of the stationary phase the column length is only 0 cm, resulting in an analysis time of less than minutes. Application 9 - Application - Mobile phase: Gradient: Temperature: Sample size: 0 µl Concentration range: µg/ml Sample solvent: mobile phase. malonamide -ethyl, -phenyl. ethosuximide. primidone. carbamazepine 0,-epoxide. phenobarbital. carbamazepine. phenytoin 0 mm x. mm, dp = µm Cat. no. acetonitrile/water 0- min acetonitrile/water = / (v/v) -0 min acetonitrile/water = 0/0 (v/v) 0- min acetonitrile/water = 0/0 (v/v).0 ml/min ambient UV, 0 nm 00 mm x mm, dp = µm Cat. no. 0 Mobile phase: methanol/0.0 M ammonium biphosphate (ph =.) = 0/0 (v/v) 0. ml/min Temperature: 0 C UV, 0 nm Sample size: 0 µl Concentration range: - 0 ppm (w/v) Sample solvent: mobile phase. uracil Conc. = ppm (w/v). protriptyline Conc. = 0 ppm (w/v). nortriptyline Conc. = ppm (w/v). doxepin Conc. = 0 ppm (w/v). imipramine Conc. = ppm (w/v). amitriptyline Conc. = ppm (w/v) 0 min 0 9 min

7 Antiseptics Application 9 - The inertness, efficiency and selectivity of the OmniSpher C column permits the fast, simultaneous determination of cetrimide and cetrimide in products in accordance with the British and European Pharmacopoeia. The determination of chlorhexidine in products give excellent validation results with good linearity and an RSD of better than 0.% on peak areas. Mobile phase: Pressure: 0 mm x. mm, dp = µm Cat. no. acetonitrile/water (+ 0. M perchloric acid + g Na Octanesulphonate) = 0/0 (v/v) 0. ml/min bar (00 psi) UV, 0 nm J. Barberio, Agroserve Ltd, Warminster, Wiltshire, UK. hexadecyltrimethylammonium bromide 9. mg/ ml. chlorhexidine 0 0 min Analgesics Several classes of drugs are analyzed with the new, universal packing material for RPLC. A mixture of most common analgesics is separated in 0 minutes, all peaks showing an efficiency above 0,000 plates for this cm column. Anticoagulantia A fast separation of -hydroxycoumarin anticoagulantia is possible under isocratic conditions using a 0 cm x.0 mm ID column. Application - Application - 0 mm x. mm, dp = µm Cat. no. Mobile phase: acetonitrile/ mm phosphate buffer (ph ) = 0/0 (v/v) ml/min Temperature: C UV, nm Sample size: µl 00 mm x mm, dp = µm Cat. no. 0 Mobile phase: acetonitrile/water (with 0.% acetic acid, ph=. with ammonia) = /0 (v/v) 0. ml/min UV, 0 nm Sample size: 0 µl Th. Welsch and A. Köhne, University of Ulm, Germany H. Thijssen, Dept. of Pharmacology, University of Maastricht, The Netherlands. acetaminophen. salicylic acid. carbamazepine. diflunisal. ketoprofen. naproxen. flurbiprofen. indomethacin. acenocoumarol. coumatetralyl. warfarin 0 min 0 0 min

8 Vitamin A Application - Several isomers of Vitamin A precursors (carotene) are analyzed in about minutes. A non-aqueous mobile phase is used to obtain the desired selectivity and retention. 0 mm x. mm, dp = µm Cat. no. Mobile phase: acetonitrile + methanol + n-hexane + triethylamine = (v/v). ml/min UV, 0 nm Sample size: 0 µl Concentration range: 0 mg/00 ml Sample solvent: acetone/chloroform = 9/ (v/v). g-carotene. trans b-carotene. -cis b-carotene. b-zeacarotene 0 0 min Steroids Using a non-buffered eluent, a mixture of corticosteroids is separated on a cm OmniSpher C column. Selectivity, column efficiency and peak shape meet the standards of with state-of-the-art column technology. Additives A very fast analysis of the preservative phenoxyethanol is possible with a 00 x.0 mm ID cartridge column. It takes about one minute to complete an analysis with a mobile phase of % methanol / phosphate buffer. Application - Application - 0 mm x. mm, dp = µm Cat. no. Mobile phase: acetonitrile/water = 0 / 0 (v/v) ml/min Temperature: C UV, nm Sample size: µl Th. Welsch and A. Köhne, University of Ulm, Germany 00 mm x mm, dp = µm Cat. no. 0 Mobile phase: methanol/sodium dihydrogen orthophosphate buffer ( g/l) = / (v/v) ml/min UV Sample size: 0 µl Concentration range: ppm Sample solvent: mobile phase. -phenoxyethanol. hydrocortisone. nortestosterone. -dehydro-a-methyltestosterone. testosterone. a-methyltestosterone. progesterone 0 0 min 0.0 min

9 Food & beverage Organic acids A range of free organic acids can be separated using a low modifier gradient of an acidified mobile phase. Small molecules (like formic acid) and the very polar component lactic acid can be elutes without special additives, such as ion paring agents. Application 0-0 mm x. mm, dp = µm Cat. no. Mobile phase: solvent A = 0.% HPO in water; solvent B = methanol Gradient: 0-0 min 00% A to 90% A: 0% B (v/v) at %/min ml/min Temperature: ambient UV, nm Sample size: µl Concentration range: µg/ml Sample solvent: solvent A. oxalic acid. glucuronic acid. formic acid. ascorbic acid. lactic acid. acetic acid. maleic acid. citric acid 9. succinic acid 0. levulinic acid min Aflatoxins After sample preparation and concentration by antibody technology, 0 µl of baby food extract is injected on a.0 mm ID OmniSpher C column. The reduced internal diameter results in a lower sample dilution in the column, hence in higher peaks in the final chromatogram (generated by the selective and sensitive fluorescence detector). Detection limits are in the ppt-range. Flavonoids Polyphenols, due to the presence of large numbers of OH-groups in the molecules, tend to give tailing peaks on many standard reversed phase columns. With the OmniSpher C column excellent symmetrical peaks are obtained with significantly better retention time stability than on other columns. Application - Application - 0 mm x.0 mm, dp = µm Cat. no. 0 Mobile phase: methanol/acetonitrile/water = 90/0/0 (v/v/v) 0. ml/min Temperature: 0 C Sample size: 0 µl Concentration range: 0-00 ng/l. aflatoxin G 0 ng/l. aflatoxin G 00 ng/l. aflatoxin B 0 ng/l. aflatoxin B 00 ng/l FLD, Fluorescence Detection lext = nm; lem = nm 0 mm x. mm, dp = µm Cat. no. Mobile phase: A = 0.% TFA in water; B = acetonitrile/0.% TFA in water (0/0) Gradient: % B (hold min) to % B in min; to % B at min 0.9 ml/min Temperature: 0 C Pressure: 00 bar UV, 0 and 0 nm Sample size: 0 µl Concentration range: mg/l Sample solvent: methanol. catechol. epicatechin. rutin. myricetin. morin. quercetin. naringenin. apigenin 9. kaempferol H.-G. Janssen and A. Louter, Unilever Research, Vlaardingen, The Netherlands 9 0 nm 0 min 9 0 min 0 nm 0 min

10 Environmental Nitrogen containing compounds A mixture of several anilines and heterocyclic amines can be separated in an isocratic run without a buffer in the mobile phase. Good peak shape is obtained for compounds such as pyridine and aniline (benzylamine). Explosives Under non-buffered conditions OmniSpher C quickly separates a mixture of amino-nitro aromatics. Sensitivity and analysis time can be improved by using a 0 cm x.0 mm ID column without other modifications. Application - Application - 0 mm x. mm, dp = µm Cat. no. Mobile phase: acetonitrile/water = 0/0 (v/v). ml/min Temperature: ambient UV, nm Sample size: µl Concentration range: 0 ppm Sample solvent: mobile phase. caffeine. pyridine. aniline. m-toluidine. o-toluidine. -aminotoluene. N-methylaniline. -ethylaniline 9. -nitroanisole 0. -nitroaniline 0 mm x. mm, dp = µm Cat. no. Mobile phase: acetonitrile/water = / (v/v) ml/min Temperature: C UV, nm Sample size: µl.,-diamino--nitrotoluene. m-dinitrotoluene. -amino--nitrotoluene. -nitroaniline. -amino-,-dinitrotoluene.,-dinitrotoluene.,,-trinitrotoluene Th. Welsch and A. Köhne, University of Ulm, Germany min 0 min Varian Analytical Instruments North America: , Europe The Netherlands: Asia Pacific Australia:.90. Latin America Brazil:..0 Other sales offices and dealers throughout the world GC GC/MS AAS ICP ICP-MS UV-Vis-NIR Fluorescence NMR Analytical Supplies

columns Acclaim Mixed-Mode WCX-1 for Separating Basic Molecules

columns Acclaim Mixed-Mode WCX- for Separating Basic Molecules The Acclaim Mixed-Mode WCX- is a novel, high-efficiency, silica-based column specially designed for separating various basic analytes. This