Passion. Power. Productivity.

Transcription

1 - - Passion. Power. Productivity.

2 Acclaim Acclaim Acclaim PA/PA Acclaim A Acclaim Acclaim PA Acclaim Explosives rdering and Warranty Information In the U.S. All rders Including: Credit Card rders (VISA, American Express, MasterCard) Blanket rders Emergency rders Call -8-Dionex- ( ) or FAX rdering by Credit Card Dionex accepts payment by MasterCard, VISA, and American Express cards with minimum orders of $.. Call -8-DINEX- ( ) for more information. Acclaim A Acclaim PA/PA Acclaim Blanket rders Dionex will arrange for blanket orders with a specified shipping time. Please call -8-DINEX- ( ) for further information. utside the U.S. Please contact your local Dionex representative. Acclaim Acclaim PA/PA Acclaim A Acclaim Surfactant Shipping and Handling rders placed for consumables are typically shipped within two working days. Dionex ships FB Sunnyvale, CA, for all U.S. orders, unless prior arrangements have been made. For UPS, Federal Express, or any carrier under contract with Dionex, Dionex will prepay the freight charges and add the charges to the invoice. Unless otherwise specified, shipments are made by ground transportation. Acclaim Acclaim Acclaim PA 7 by Dionex Corporation All rights reserved LPN 668- Dionex Corporation 8 Titan Way P.. Box 6 Sunnyvale, CA AQA and MSQ are trademarks of Thermo Electron Corporation. ne-a-day is a registered trademark of Bayer Corporation. NyQuil is a registered trademark of Proctor and Gamble. All other trademarks and registered trademarks are property of Dionex Corporation. - -

3 Contents PART NE Introduction... Acclaim Column Familes at a Glance... Selecting an Acclaim Column...6 Understanding More about the Bonded Phase... 8 Mobile Phase Considerations...9 Acclaim Quality... PART TW Chapter : Acclaim C8... Chapter : Acclaim C8... Chapter : Acclaim C8...6 Chapter : Acclaim Polar Advantage (PA)...8 Chapter : Acclaim Polar Advantage II (PA)... Chapter 6: Acclaim A for rganic Acids... Chapter 7: Acclaim Surfactant... Chapter 8: Acclaim Explosives E and E...6 PART THREE Applications... 8 Applications Contents...8 Pharmaceutical Applications...9 Environmental Applications... Food and Beverage Applications...7 Chemical (Surfactant) Applications... Bioscience Applications...6 Natural Products Applications...8 rganic Acid...9 PART FUR Appendix... Lot Qualification Tests... Topical Index... Part Number Index... Analyte Index

4 Introduction INTRDUCTIN Par t I: In t r o d u c t i o n Acclaim Column Families at a Glance Dionex Acclaim columns are silica-based columns designed for high efficiency separations and manufactured using ultra-high purity silica. Product families consists of a broad spectrum of stationary phases in the form of the conventional C8 and C8 phases. polar embedded phases. and several specialty phases, each designed for a specific application or group of applications. All Acclaim columns are designed and manufactured at Dionex, to tight specifications, to ensure consistent performance. General Purpose HPLC Columns Acclaim High purity silica for small molecule separations Acclaim reversed-phase C8 and C8 phases are manufactured using high purity silica with a -Å pore diameter, very high surface coverage, low silanol activity, and very low metal content. All the phases are LC-MS compatible with good capacity ideal for use with high organic mobile phases. Characteristics High purity spherical silica, LC/MS compatible, monomeric bonding, endcapped, Å pore size, µm, µm. Acclaim Wide pore columns for macromolecule applications Acclaim columns are ideal for the reversed-phase separation of proteins, peptides, and other biological macromolecules. The unique bonding chemistry results in a high-density, highly uniform phase coverage. The use of a -µm silica particle results in fast, high-resolution separations. Characteristics High purity spherical silica, LC/MS compatible, monomeric bonding, endcapped, Å pore size, µm, µm. Acclaim PA Polar-embedded Reversed-phase columns for separating a wide range of analytes Acclaim PolarAdvantage (PA) columns feature a patented bonding chemistry that incorporates a polar sulfonamide group near the surface of the silica particle. This unique chemistry provides enhanced hydrolytic stability and allows a single column to resolve both polar and nonpolar analytes. The Acclaim PA is compatible with % aqueous mobile phases and exhibits high reversed-phase capacity. Characteristics High purity spherical silica, LC/MS compatible within guidelines, endcapped, Å pore size, µm, µm. Acclaim PA Polar-embedded columns with enhanced hydrolytic stability Acclaim PolarAdvantage II (PA) columns feature a patented bonding chemistry that provides enhanced hydrolytic stability from ph. and allows a single column to resolve both polar and nonpolar analytes. The Acclaim PA is compatible with % aqueous mobile phases and exhibits high reversed-phase capacity, with selectivity complementary to conventional C8 columns such as the Acclaim C8. Characteristics High purity spherical silica, LC/MS compatible, endcapped, Å pore size, µm, µm. - -

5 Introduction Acclaim General Purpose Column Specifications Dionex phase Acclaim C8 Acclaim C8 Acclaim C8 Acclaim PA Acclaim PA Bonded phase C8 C8 C8 Sulfamido C6 Amide C8 USP type L L7 L na na Endcapped Yes Yes Yes Yes Yes Substrate Ultrapure silica Ultrapure silica Ultrapure silica Ultrapure silica Ultrapure silica Particle shape Spherical Spherical Spherical Spherical Spherical Particle size and µm and µm µm and µm and µm Metal impurity (ppm) < ppm < ppm < ppm < ppm < ppm Na, Fe, Al Average pore diameter Å Å Å Å Å Surface area (m /g) Total carbon content 8% % 7% 7% 7% INTRDUCTIN Specialty HPLC Columns Acclaim A Specialty column for fast organic acid analysis The Acclaim A column features a patented polar-embedded stationary phase optimized and use tested for hydrophilic organic acid separations. The Acclaim A stationary phase is compatible with % aqueous mobile phases and has excellent hydrolytic stability at low phs. Characteristics High purity spherical silica, LC/MS compatible, monomeric bonding, endcapped, Å pore size µm, µm. Acclaim Surfactant Specialty column for the separation of surfactants The Acclaim Surfactant column uses high purity -Å silica and a proprietary bonded phase to separate anionic, nonionic, cationic and amphoteric surfactants in various matrices, including consumer products, pharmaceutical formulations and environmental samples. Characteristics High purity spherical silica, monomeric bonding, endcapped, Å pore size. Acclaim Explosives Specialty columns for the separation of explosives. A total solution to EPA Method 8; two columns each of which provides guaranteed baseline resolution of all explosives targeted by the EPA. With their complementary selectivities, the E and E provide both primary and confirmatory columns. Characteristics High purity spherical silica, LC/MS compatible, endcapped, Å pore size Acclaim Specialty Column Specifications Dionex phase Acclaim A Acclaim Surfactant Acclaim Explosives E Acclaim Explosives E Bonded phase Proprietary Proprietary Proprietary Proprietary USP type na na na na Endcapped Yes Yes Yes Yes Substrate Ultrapure silica Ultrapure silica Ultrapure silica Ultrapure silica Particle shape Spherical Spherical Spherical Spherical Particle size µm µm µm µm Metal impurity (ppm) < ppm < ppm < ppm < ppm Na, Fe, Al Average pore diameter Å Å Å Å Surface area (m /g) Total carbon content 7% na na na - -

6 Introduction INTRDUCTIN Selecting an Acclaim Column There are four questions to consider when choosing an HPLC column:. What are the goals of the separation? Speed of analysis Critical peaks requiring resolution Solvent consumption/minimal waste Sensitivity requirements perating costs ther criteria. What are the best silica characteristics for this separation? This is explained in the section below.. What is the most appropriate column format? see page 8 for a summary of the impact of column dimensions on the separation. What is an appropriate stationary phase chemistry? see pages for a summary of the Acclaim family of columns and Part for detailed information. Choosing the Best Column Chemistry When a chromatographer designs a separation, selectivity of the stationary phase is the first of many factors that must be considered. Selectivity is the result of the differing interactions between each analyte and the stationary phase. Determining the column with optimal selectivity is the essential starting point. This determination depends on the nature of the desired separation. The easiest first step in choosing a column is to identify whether there is a column designed specifically for your separation. This can be readily established if the column is named according to its application, as in the case of some of the Dionex Acclaim columns. Thus, if you are separating surfactants, the Acclaim Surfactant column is the best first choice; likewise Acclaim Explosives column are used for EPA Method 8, and Acclaim A is for organic acid analysis. If there is no specialty column for your application, then you will need some understanding of chemistry of your sample to choose the best column chemistry. For those cases where the nature of the sample is completely unknown, the best first choice is usually C8 because of its excellent peak efficiency, low silanol activity, and ability to separate many organic molecules. The C8 phase is a good choice if you want less retention of the sample on the column. If you need highly aqueous mobile phase conditions, or different selectivity, then the polar embedded phases of the Acclaim Polar Advantage are the best option. For samples that are well characterized, various tests are useful to characterize the stationary phases and to compare different columns. The tests include studies of effects such as hydrophobic interactions, molecular shape, and ion-exchange interactions between the stationary phase and the sample analytes. A good, purely reversed-phase interaction will exhibit strong retention of strongly hydrophobic molecules and this will be the only retention mechanism. As the density of the bonded phase decreases, other retention mechanisms will come into play, such as ion exchange interactions between the sample and residual silanol groups on the surface of the silica gel, or sieving effects for similar sized molecules of different shapes. Thus, these studies can be used to choose a column with specific characteristics, or to find a new column with characteristics similar to an old favorite. Hydrophobicity The strength of the hydrophobic interaction between an analyte and the bonded silica is usually characterized by measuring the selectivity of ethylbenzene relative to toluene. Because of their high-density bonding, Acclaim C8 and C8 columns have near maximum hydrophobicity compared with other C8 and C8 columns, and therefore almost pure reversedphase behavior. Ion-Exchange Activity Ion-exchange activity is usually present where low surface coverage of the bonded phase, or insufficient endcapping is present. Since this effect is generally not manufactured into reversed phase stationary phases on purpose, ion-exchange activity of reversed phase columns is not reproducible and separations reliant on this effect are not reliable. Selectivity between phenol, aniline and pyridine is affected by the degree of their ion-exchange activity with residual surface silanol groups at ph 7, because under these conditions the two bases are protonated but phenol is neutral. As illustrated in Acclaim Selectivity figure, overall the Acclaim columns exhibit low ion-exchange activity under neutral conditions and the general purpose C8 and C8 show particularly low ion-exchange activity, emphasizing their high surface coverage of bonded phase. Molecular Shape When hydrophobic interaction alone is insufficient to produce the required separation, it is often convenient to have secondary effects that will have an impact. Triphenylene and o-terphenyl are both tricyclic hydrocarbons of similar size but with different shapes. The type of bonding chemistry of the stationary phase and the bonding density will both influence how these compounds interact with a stationary phase. These compounds are unresolved on the C8 phase, similarly resolved on the -Å or -Å C8 phases but as shown in the figure above, shape differences have a much larger effect with the polar embedded phases than they do with C8 phases. Hydrogen Bonding and Dipole-dipole Interactions In addition to hydrophobic retention, and ion exchange if the stationary phase has exposed charged sites, phenol, aniline and pyridine are also retained by dipole-dipole interactions and hydrogen bonding. These secondary effects can be exploited to improve a separation by choosing a stationary phase that has stronger interactions of this type, such as the polar embedded phases

7 Introduction INTRDUCTIN Acclaim Selectivity Column: Mobile Phase: Temperature: Flow Rate:..6 mm 9/ w/w methanol/phosphate buffer, ph 7. C ml/min Column Chemistry PA C8 C8 C8 Column: Mobile Phase: Temperature: Flow Rate: PA 9.8 Column Chemistry C8 C8 C8 PA PA 8 Ethylbenzene Toluene.8 Ethyl benzoate Dimethylaniline.6 Nitrobenzene. Phenol Aniline Pyridine.. Polar Compounds Retention Time Relative to Toluene Relative Retention Time vs Toluene.6 mm 9/ v/v methanol/water C ml/min 6 Triphenylene o-terphenyl Ethylbenzene Toluene Hydrophobic Compounds Probe Molecules Property Indicator Ethylbenzene/toluene Hydrophobic interaction Triphenylene and o-terphenyl Selectivity for molecular shape Phenol, aniline and pyridine Dipole-dipole interactions and hydrogen bonding Aniline and pyridine Ion exchange 87 Choosing the Appropriate Particle Size Choosing the Pore Size and Surface Area Particle size affects peak efficiency, which in turn impacts resolution or the amount of space between peaks. Smaller particles sizes give higher efficiencies and higher resolution than larger ones. However, at the same flow rate, the backpressure will be greater using smaller particle sizes than larger ones. Therefore, smaller particle sizes are recommended for fast analyses, using shorter columns. Select a pore size appropriate for the molecular weight of your sample analytes. The pore size should be large enough to allow the sample molecules of interest to enter and pass through. If the pore size is too small, size exclusion effects can cause unwanted peak broadening. Acclaim columns are available packed with either -µm or -µm bonded silica. A column packed with -µm material will give almost twice the separation efficiency as a column of identical proportions, packed with -µm material, but the backpressure will be four times as high on the -µm column at the same linear velocity. µm MW < kda: use Acclaim, Acclaim PA or Acclaim PA MW < kda: use Acclaim Generally speaking, the smaller the pore size, the greater the number of pores and the higher the surface area. The higher the surface area, the higher the capacity of the packing material. This means that the retention time of a given analyte will be shorter on a wide-pore material than on a narrow-pore one. µm Smaller pores = Larger surface area Larger pores = Smaller surface area -7-

8 Introduction INTRDUCTIN Understanding More about the Bonded Phase Dionex offers highest quality silica-based reversed phase packing materials. The type of bonding, carbon load and endcapping are all factors that contribute to the superior bonded phase. Type of Bonding Functional groups are attached to the silica substrate either via a single attachment (monomeric bonding), or via multiple attachments (multidentate bonding). Monomeric bonded phases provide higher column efficiencies, than polymeric phases, but polymeric phases are very stable under ph extremes. Carbon Load and Endcapping The Percent (%) carbon is a rough guide to the capacity of the column. Given the same type of silica the higher carbon load is an indicator of surface coverage. Phases with higher carbon loads are more strongly hydrophobic, resulting in higher capacity, longer retention times, and often better resolution. When the functional groups are attached to the silica particle, not all silanol groups on the surface of the silica particle are covered. Free silanol groups will interact with polar analytes, altering the retention times and often causing peak tailing for organic bases. To minimize these secondary interactions, the free silanol groups are endcapped. All Acclaim column packings are endcapped. Choosing the ptimum Column Format Column efficiency, which is measured in theoretical plates (N), is mostly a function of the column length and particle size. Acclaim columns are available in many formats, therefore it is important to carefully select the best one for a particular application. Column Length Shorter columns provide faster run times and longer columns provide better resolution. If all else is constant, peaks are better separated on longer columns. However, the pressure, mobile phase consumption and time of analysis all increase in proportion to column length, thereby raising the cost per analysis. Detection limits are better on shorter columns. Column Length Column Length Selection Application 7 mm Fast analysis. Works best with -µm particles size mm Standard separations. Works well with - or -µm particle sizes > mm High resolution applications. Works best with -µm particle size Column Diameter Select a column diameter according to your requirements for sample size, solvent consumption and sensitivity. Column diameter impacts sensitivity (narrow columns provide better sensitivity), loading capacity (wider is better for larger sample volumes and for purifying samples), solvent consumption (narrow columns use less solvent) and backpressure (narrow columns exhibit higher backpressure so you will need to decrease the flowrate). Scale the flow rate by the square of the column diameter to preserve backpressure and retention time. To gain the full benefits of smaller column diameters, your HPLC instrumentation needs to be optimized for the flow rate. Column Diameter Column Diameter Selection Application.7 to. mm I.D High sensitivity or limited volume samples. Use with LC/MS instrumentation. mm I.D High sensitivity or limited volume samples. Use with low dead-volume instrumentation. mm I.D Lower solvent consumption. Use with standard instrumentation..6 mm I.D Standard formats. Use with standard instrumentation Quick Tips when Choosing Column Format ptimize the analysis time and chromatographic efficiency by evaluating the trade-offs between the particle diameter and column lengths. As a general rule, the maximum sample load, solvent consumption, and detection limits are proportional to the square of the column diameter. Scale the flow rate by the square of the column diameter to preserve backpressure and retention time: a.-mm column requires % of the flow rate of a.6-mm column. Given the same mass of sample, the peak area scales according to the inverse square of column diameter; for example, a.-ng sample injected on a.-mm column would yield.8 times more area than on a.6-mm column. A -µm,.6 -mm column delivers similar peak widths as a -µm,.6 -mm column in 6% of the time. -mm columns sometimes provide adequate resolution while saving time and solvent. LC-MS applications can increase throughput using shorter column or larger particle size than required for UV detection. When using gradient elution, to preserve the relative elution times, scale the gradient time proportionally to the column length. When run time is more important than sharp peaks, optimize the resolution by shortening the column instead of lengthening the gradient. By default, a -µm, -mm-long column is always a good starting point

9 Introduction Mobile Phase Considerations The effect of mobile phase composition on column lifetime should be part of the consideration for designing a method. Use the highest practical quality of water, solvent, and buffer components; HPLC-grade material has low UV absorbance and is submicron-filtered by the manufacturer. To prevent fouling of your Acclaim column, use mobile phases that have been filtered through a.-µm or smaller filter. Both alkaline conditions and strong acid conditions will degrade silica-based columns over time. The silica substrate resists acids, but is soluble in alkali. The upper ph limit is determined by the rate of dissolution of the silica gel, which is also affected by buffer ions and organic co-solvents. When organic solvents are added to buffers of inorganic anions such as phosphate, the buffer becomes more alkaline; when organic solvents are added to buffers of organic bases, such as TRIS, the buffer becomes less alkaline Acid conditions hydrolyze the bonded layer, rather than the silica itself, and the rate of this effect is proportional to the hydrogen ion concentration. The rate of hydrolysis is slower toward the middle of the ph range. Fortunately, organic modifiers usually provide protection against degradation. INTRDUCTIN - 9 -

10 Introduction INTRDUCTIN Acclaim Columns Dedicated to High Quality Reliability and Durability At Dionex we know that quality and reliability are essential to a successful analysis. ur columns are thoroughly tested individually, so that chromatographers can have full confidence in them. Manufacturing starts with an ultrapure silica substrate, using only carefully selected lots with narrow ranges of physical parameters. By design, the bonding processes are clean and repeatable with no unexpected changes in performance. Each batch of bonded silica receives a full suite of validation tests appropriate to its intended use. The bonded silica is packed in precision-polished 6 stainless steel hardware using highly reliable processes. Each packed column is tested to ensure the same great performance every time. The quality assurance reports for silica lot validation and column performance explain the test protocols, list the specifications, and show the actual chromatograms. Base Asymmetry... Acclaim Columns Base Performance: Amitriptyline D C B A Amitriptyline Performance: # Acclaim Type EP Asym. N/m D Å, µm, C8. 8K C Å, µm, C8. 6K B Å, µm, PA C6. K A Å, µm, C8. K. Uracil. Propranolol. Toluene. Doxepin. Amitriptyline 8 Performance Indicators Acclaim bonded-silica columns have been designed to meet the high quality standard needed in laboratories today. The innovative surface chemistries deliver exceptional peak efficiencies for a broad range of analytes. To ensure optimal performance, all Dionex Acclaim products are thoroughly characterized using a number of performance indicators, including surface coverage of the bonded phase, metal contamination, steric selectivity, column polarity, column hydrophobicity, and low silanol activity for bases. The specialty columns are also application-tested for their specific analysis, to ensure that each lot of bonded silica provides high-performance separations. The following charts and chromatograms explain how to interpret each of these tests. Good surface coverage of the silica gel with the bonded phase, and exhaustive endcapping ensure that Acclaim columns provide symmetrical peaks and superior performance for the separation of basic drugs. Amitriptyline is a well-known example of a basic pharmaceutical that interacts strongly with residual silanol groups on the silica gel, to produce asymmetrical peaks. Acid Asymmetry Peak Asymmetry of Acids Polarity Index Polarity = [ K BBA K phenol /K toluene] Polar-Embedded Leading Brand E (As. =.) (As. =.9) Acclaim PolarAdvantage Mobile Phase: /6 v/v mm phosphate buffer, ph./ch CN Flow Rate:. ml/min Inj. Volume: µl Temperature: C Detection: nm. Uracil. Pyridine. Phenol. N,N-Dimethylaniline. p-butylbenzoic acid 6. Toluene 76 Acclaim C8 Luna() C8 Acclaim PA SymmetryShield C8 Discovery RP-Amide C6 ProntoSIL EPS C8 Acclaim PA ZRBAX Bonus-RP Many polar-embedded columns perform well for a limited group of compounds. Acclaim PolarAdvantage columns use state of the art methods of synthesis to eliminate unwanted secondary interactions. The result is outstanding performance for hydrophilic and hydrophobic acidic, basic, and neutral analytes. The polarity index chart above ranks commercially available stationary phases according to their polarity relative to one another, under the same conditions. The more similar the polarity index of two stationary phases, the more similar their separation of polar compounds should be. If one column does not provide adequate resolution, choosing a column with a different polarity index may deliver a better separation. - -

11 Introduction Metal Contamination Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: / v/v water/methanol Temperature: C Flow Rate:. ml/min RA.7 Evaluation of Metal Contamination Inj. Volume: µl Detection: nm Samples:.,'-bipyridyl ng/µl.,'-bipyridyl Competitor A, µm Steric Selectivity Comparison of Steric Selectivity for Acclaim C8 and Acclaim PA Acclaim C8 Competitor Competitor Columns: As indicated Mobile Phase: 9% methanol Temperature: C Flow Rate: ml/min Inj. Volume: µl. Toluene. Phenanthrene. o-terphenyl. Triphenylene INTRDUCTIN m Competitor B, µm Competitor C, µm Competitor D, µm Competitor E, µm Acclaim C8, µm 6 Note: Ratio of asymmetries (RA) is quoted as the asymmetry of Peak divided by Peak. Metal contamination can interfere with the separation of many different substances, resulting in peak tailing and concomitant poor peak resolution., -bipyridyl is a strong chelating agent, and will reveal any contamination by metal ions. The figure above shows that Acclaim outperforms five competitor columns in this test , Competitor Competitor Competitor Acclaim PolarAdvantage 6 8. Toluene. Phenanthrene. o Terphenyl. Triphenylene Me 77 Column: µm Dimensions:.6 mm Mobile Phase: 9/ v/v MeH/D.I. H Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: UV, nm Depending on the bonding chemistry, various C8 columns can have different selectivity to molecular shape. Acclaim may be just what you need for your unique separation challenge

12 Acclaim C8 C8 Part Two: Columns Chapter : Acclaim C8 High performance reversed-phase columns for the separation of small molecules The Acclaim C8 series columns feature a densely bonded monolayer of octadecyldimethylsiloxane on a highly pure, spherical, silica substrate with Å pore structure. The Acclaim C8 columns are the classic reversed-phase columns. These columns are recommended for general-purpose reversed-phase applications where high surface coverage, low silanol activity, and excellent efficiency are required. The Acclaim C8 columns feature: Highly efficient, symmetrical peaks for difficult basic and chelating analytes Ultrapure silica substrate ptimized surface pretreatment, proprietary highdensity bonding process, and double endcapping Reliability designed into the manufacturing process and assured by thorough and appropriate testing High hydrophobicity and low polarity yield high selectivity for hydrophobic substances LC/MS compatible Wide range of applications in pharmaceutical, environmental, food testing, and product-quality testing for small molecules Bonding USP code Endcapping C8 monomeric L Double % C Pore size CH Si CH (CH ) CH CH 6 Physical Specifications Å Surface area 9 m /g Particle diameter.9. or.. µm Performance Specifications Lot Polar selectivity ratio..6 Base asymmetry.98. ( µm).98.6 ( µm) Metal activity ratio.9. Steric selectivity ratio.8.6 Column Efficiency (N/m) 9, ( µm), ( µm) Asymmetry (EP).98. Retention time ( µm) ( µm) Chromatographic specifications for Acclaim C8.6 mm columns. Please refer to Part for a detailed description of the tests and specifications. Specifications are subject to change at the discretion of Dionex b 8--6a Control Chart for Acclaim C8 µm 6 7 k' Control chart showing k (phenanthrene) for nine lots of Acclaim -µm C8. The data show the average and standard deviation for all columns shipped from that lot. LC-MS of Pesticides 8 Column: Acclaim C8, µm Dimensions:. mm Mobile Phase: 7/% methanol/.% formic acid Flow rate. ml/min Detection: AQA MS, +ESI, V, C, SIM as indicated m/z mass injected. Monuron 99 ng. Altrazine 6. ng. Propazine ng. Diuron. ng. Linuron 9 ng Recommended Ranges of peration To maintain wetting of the hydrophobic surface, the recommended range for organic composition is %. The type 6 stainless steel column hardware is compatible with common HPLC mobile phases. To prevent corrosion, high concentrations of halide ions at ph <. should be avoided. ph Range. 8. Pressure limit psi Temperature < 8 C

13 Acclaim C8 LC/MS Compatibility Acclaim C8 columns are LC/MS compatible. LC/MS analysis shows very low bleed for this column. ut of the box, this column is ready to use after only a few minutes of conditioning with solvent. The same low bleed is attained over the entire ph range of 8. The baseline obtained from an Acclaim column is very similar to that obtained from an empty capillary tube. Column Bleed: Acclaim C8, ph 8 Mobile Phase: (A) Ammonium bicarbonate, ph 8 (B) Acetonitrile Gradient: 8/ to / at min, to 8/ at min MS: Positive ion electrospray, full scan 6 m/z,, TIC,, TIC Å C8, µm No column Counts Counts C8,, Counts Counts ,, Counts amu Counts amu 9 rdering Information: Acclaim C8 Standard particle sizes are nominally and µm. Analytical columns are available in.- and.6-mm diameters; standard lengths are,, and mm for - and -µm particles and mm for -µm particles. Capillary formats are available in 7 and µm,.-mm and custom diameters, and in -, -, or -mm lengths. Micro precolumn and nano precolumn cartridges are available in several formats. Guard columns in both.- and.-mm sizes packed with -µm particles are recommended to protect both the and µm analytical columns. Please order through your local Dionex office or distributor. Refer to the following part numbers. Acclaim C8 Analytical Columns µm. mm µm. mm µm. mm... 9 µm.6 mm... 9 µm.6 mm... 9 µm.6 mm... 9 µm. mm... 9 µm. mm... 9 µm. mm... 9 µm. mm... 9 µm.6 mm µm.6 mm µm.6 mm µm.6 mm Acclaim C8 Guard Columns µm. mm µm. mm Biocompatible cartridge holder for. and. mm guards...96 Guard to analytical column coupler...97 Holder and coupler kit Acclaim C8 Micro and Nano Columns µm 7 µm mm (NAN7) µm 7 µm mm (NAN7) µm µm mm (FUS) µm µm mm (FUS) µm mm (MIC) µm mm (MIC) µm 7 µm mm (NAN7) µm 7 µm mm (NAN7) µm 7 µm mm (NAN7) µm µm mm (FUS) µm µm mm (FUS) µm µm mm (FUS) µm mm (MIC) µm mm (MIC) µm mm (MIC) Acclaim C8 Micro and Nano Precolumns µm µm mm (pack of )...66 µm µm mm (pack of )...66 µm µm mm (pack of )...6 µm µm mm (pack of )...6 µm 8 µm mm (pack of )...6 µm mm (pack of )....6 µm mm (pack of )....6 These columns are designed for optimal performance using Dionex UltiMate and ICS- chromatography instruments. - -

14 Acclaim C8 C8 Chapter : Acclaim C8 High performance reversed-phase columns for the separation of small molecules CH Si CH (CH ) CH CH 6 The Acclaim C8 series columns feature a densely bonded monolayer of octyldimethylsiloxane on a highly pure, spherical silica substrate with a Å pore structure. The Acclaim C8 series employs the same bonding chemistry and substrate as C8, and therefore features the same high standards of efficiency, coverage, and silanol activity. The Acclaim C8 features: Highly efficient, symmetrical peaks with difficult basic and chelating analytes Ultrapure silica substrate ptimized surface pretreatment, proprietary highdensity bonding process, and vigorous endcapping Reliability designed into the manufacturing process and assured by thorough and appropriate testing Less hydrophobic, less retentive than C8 LC/MS compatible Excellent performance for basic pharmaceuticals and environmental samples Performance Specifications Lot Polar selectivity ratio..6 Base asymmetry.98. Metal activity ratio.9. Column Efficiency (N/m) 9, ( µm), ( µm) Lot # Asymmetry (EP).98. Retention time.9.9 ( µm)..9 ( µm) Chromatographic specifications for Acclaim C8.6 mm columns. Please refer to Part for a detailed description of the tests and specifications. Specifications are subject to change at the discretion of Dionex Control Chart for Acclaim C8, µm... k' Control chart showing k (phenanthrene) for five lots of -µm C8. The data show the average and standard deviation for all columns shipped from that lot. 9 Bonding USP code Endcapping C8 monomeric L7 Yes % C..8 Pore size Physical Specifications Å Surface area 9 m /g Particle diameter.9. or.. µm Recommended Ranges of peration To maintain wetting of the hydrophobic surface, the recommended range for organic composition is %. The type 6 stainless steel column hardware is compatible with common HPLC mobile phases. To prevent corrosion, high concentrations of halide ions at ph <. should be avoided. ph Range. 8. Pressure limit psi Temperature < 8 C Beta-Blockers Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: /9 w/w MeH/ mm phosphate, ph 7. Flow Rate:. ml/min Temperature: C Inj. Volume: µl Detection: UV, nm. Maleic acid. Acebutolol µg/ml. Metoprolol. Timolol. xprenolol 6, 7. Labetalol diasteromers 8. Propranolol Acclaim C8 has excellent performance for basic drugs such as these beta-blockers. Peaks are efficient and symmetrical

15 Acclaim C8 LC/MS Compatibility Acclaim C8 columns are LC/MS compatible. LC/MS analysis shows very low bleed for this column. ut of the box, this column is ready to use after only a few minutes of conditioning with solvent. The same low bleed is attained over the entire ph range of 8. The baseline obtained for an Acclaim column is very similar to that obtained for an empty capillary tube. Column Bleed: Acclaim C8, ph 8 Mobile Phase: (A) Ammonium bicarbonate, ph 8 (B) Acetonitrile Gradient: 8/ v/v to : at min, to 8/ v/v at min MS: Positive ion electrospray, full scan 6 m/z,, TIC,, TIC Å C8, µm No column Counts Counts, Counts , Counts C8,, Counts Counts amu amu 9 rdering Information: Acclaim C8 Standard particle sizes are nominally and µm. Analytical columns are available in.- and.6-mm diameters; standard lengths are,, and mm for - and -µm particles plus mm for -µm particles. Capillary formats are available in 7 and µm,.-mm and custom diameters, and in -, -, or -mm lengths. Micro precolumn and nano precolumn cartridges are available in several formats. Guard cartridges in both.- and.-mm diameters packed with -µm particles are recommended to protect both the µm and µm analytical columns. Please order through your local Dionex office or distributor. Refer to the following part numbers. Acclaim C8 Analytical Columns µm. mm... 9 µm. mm... 9 µm. mm... 9 µm.6 mm... 9 µm.6 mm µm.6 mm µm. mm... 9 µm. mm... 9 µm. mm µm. mm µm.6 mm µm.6 mm µm.6 mm... 9 µm.6 mm... 9 Acclaim C8 Guard Columns µm. mm µm. mm Biocompatible cartridge holder for. and. mm guards...96 Guard to analytical column coupler...97 Holder and coupler kit Acclaim C8 Micro and Nano Columns µm 7 µm mm (NAN7) µm 7 µm mm (NAN7) µm µm mm (FUS) µm µm mm (FUS) µm mm (MIC) µm mm (MIC) µm 7 µm mm (NAN7) µm 7 µm mm (NAN7) µm 7 µm mm (NAN7) µm µm mm (FUS) µm µm mm (FUS) µm µm mm (FUS) µm mm (MIC) µm mm (MIC) µm mm (MIC) Acclaim C8 Micro and Nano Precolumns µm µm mm (pack of )...6 µm µm mm (pack of ) µm µm mm (pack of )...66 µm µm mm (pack of )...66 µm 8 µm mm (pack of )...66 µm mm (pack of ) µm mm (pack of ) These columns are designed for optimal performance using Dionex UltiMate and ICS- chromatography instruments. - -

16 Acclaim C8 C8 Chapter : Acclaim C8 Reversed-phase columns for the separation of proteins, peptides, and other biological macromolecules The Acclaim C8 series columns feature a densely bonded monolayer of octadecyldimethylsiloxane on a highly pure, spherical silica substrate with a wider, Å pore structure. The Acclaim series columns are designed for peptide mapping and separation of small proteins up to kda. The Acclaim is also useful for general-purpose, reversed-phase chromatography of small molecules. The Acclaim C8 columns feature: Technology designed for high-resolution peptide mapping applications and protein separations High-efficiency -µm spherical silica substrate The same high-performance bonding chemistry as the Acclaim series, but using a silica substrate with larger -Å pores and lower surface area Application tested for suitability in peptide mapping Minimal secondary interactions for repeatable results day-to-day and column-to-column The unique bonding chemistry results in a high-density, highly uniform phase coverage with extensive endcapping. The use of a -µm silica particle accelerates the diffusion of the mobile phase into the stationary phase, resulting in fast, high-resolution separations. Compared to -µm column packings, a given separation can be achieved in a shorter run time by increasing the flow rate of the mobile phase and running shallower gradients on shorter columns. Bonding USP code Endcapping C8 monomeric L Double % C Pore size CH Si CH (CH ) CH CH 6 Physical Specifications 6 Å Surface area 9 m /g Particle diameters.9. µm Performance Specifications Lot Base asymmetry.98. Metal activity ratio.8. Cytochrome c tryptic digest peptide map Column Efficiency (N/m), P-P:.8. min P7-P6:.6. P9-P8:.. Asymmetry (EP).98. Retention time..8 Example chromatographic specifications for Acclaim C8.6 mm columns. Please refer to Part for a detailed description of the tests and specifications. Recommended Ranges of peration The type 6 stainless steel column hardware is compatible with common HPLC mobile phases. To prevent corrosion, high concentrations of halide ions at ph <. should be avoided. ph Range. 8. Pressure limit..... psi rganic solvents % Temperature < 8 C Cytochrome c Tryptic Digest Mobile Phase: (A) 9/ water/acetonitrile,.% trifluoroacetic acid; (B) 9/ acetonitrile/water,.% trifluoroacetic acid Gradient: % A/% B to 6% A/% B at min Flow Rate:. ml/min Temperature: C Inj. Volume: µl Detection: UV, nm Chromatogram of cytochrome c tryptic digest taken from the certificate for lot #7--. Dionex monitors and controls the retention time difference between three pairs of peptides: P-P, P7-P6, and P9-P

17 Acclaim C8 Retention Time (min) Stability Study for -µm C8, Å at C P9-P8 P7-P6 P-P 6 8 Injection Number (Hours) Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: (A) 9//. (v/v/v) water/acetonitrile/tfa (B) 9//. (v/v/v) acetonitrile/water/tfa Gradient: Time (min) % A % B Temperature: C Flow Rate:. ml/min Inj. Volume: µl Inj. Amount:. µg/µl Detection: UV, nm Samples: Tryptic digest of cytrochrome c (retention data plotted for the two least separated pairs of peptides, P-P, P7-P6, and P9-P8) 79 LC/MS Compatibility Acclaim C8 columns are LC/MS compatible. LC/MS testing shows very low bleed for this column. ut of the box, this column is ready to use after only a few minutes of conditioning with solvent. Low bleed is attained over the entire ph range of 8. The baseline obtained for an Acclaim column is very similar to that obtained for an empty capillary tube. Column Bleed: Acclaim C8, ph 8 Mobile Phase: (A) Ammonium bicarbonate, ph 8 (B) Acetonitrile Gradient: 8/ v/v to / at min, to 8/ v/v at min MS: Positive ion electrospray, full scan 6 m/z,, TIC,, Å C8, µm No column TIC Counts Counts,, C8 Counts Counts ,, Counts amu Counts amu 9 rdering Information: Acclaim C8 The standard particle size for the Acclaim C8 series columns is nominally µm. Analytical columns are available in.- and.6-mm diameters; standard lengths are and mm. Guard cartridges in both.- and.-mm sizes packed with -µm particles are recommended to protect your analytical columns. Capillary formats are available in 7 and µm,.-mm and custom diameters, and in - or -mm lengths. Please order through your local Dionex office or distributor. Refer to the following part numbers. Acclaim C8 Analytical Columns µm. mm µm. mm µm.6 mm µm.6 mm Acclaim C8 Guard Columns µm. mm µm. mm Biocompatible cartridge holder for. and. mm guards...96 Guard to analytical column coupler...97 Holder and coupler kit Acclaim C8 Micro and Nano Columns µm 7 µm mm (NAN7)... 6 µm 7 µm mm (NAN7) µm µm mm (FUS) µm µm mm (FUS) µm mm (MIC) µm mm (MIC) These columns are designed for optimal performance using Dionex UltiMate and ICS- chromatography instruments

18 Acclaim PolarAdvantage PA Chapter : Acclaim PolarAdvantage (PA) Polar Embedded Reversed-Phase Columns for separating polar compounds CH CH Si (CH ) (CH ) N S CH (CH ) CH The Acclaim PA column has a patented surface chemistry that renders it compatible with solvent-free mobile phases. The ether and sulfonamide linkages are more hydrolytically stable than the amides used in many polar-embedded phases. The synthesis procedure minimize both residual silanols and amines, thus making Acclaim PA suitable for acidic, basic, or neutral analytes. Acclaim PA columns are reversed-phase silica columns with a polar-enhanced stationary phase for operation over a wider range of chromatographic conditions and with a broader application range when compared to conventional reversed-phase columns. Acclaim PA has selectivity similar to C8 for many analytes of low polarity, with the added advantage of compatibility with aqueous-only mobile phases. Some classes of compounds (for example, nitroaromatics) show significantly different selectivity patterns on this bonded phase. The high-density bonding provides good retention of hydrophilic analytes. Above 9% organic solvent composition of the mobile phase, this column starts to show some normal-phase HPLC characteristics. The Acclaim PA benefits include: Compatibility with solvent-free applications without any compromise to performance for acids and bases Novel polar-embedded surface layer Ability to work with % aqueous or % organic solvent mobile phases Resolves hydrophilic compounds High selectivity for hydrophobic compounds Different selectivity than C8 makes PA useful as a confirmation column Wide range of applications in pharmaceutical, environmental, food testing, and product-quality testing Conventional C8 Recommended Ranges of peration The type 6 stainless steel column hardware is compatible with common HPLC mobile phases. To prevent corrosion, high concentrations of halide ions at ph <. should be avoided. ph Range. 8. Pressure limit Resistance to Dewetting Before stop flow test tr = 9. min As =. Eff. = 987 plates/m After stop flow test (after cycle) tr =.9 min As =. Eff. = plates/m Acclaim PolarAdvantage Before stop flow test tr = 6. min As =. Eff. = 8967 plates/m Resistance to Dewetting After stop flow test (after cycles) tr = 6.8 min As =. Eff. = 8 plates/m psi rganic solvents % The surface of a conventional C8 phase is very hydrophobic. When hydrophobic surfaces are in contact with highly aqueous mobile phases, the partial pressure of dissolved gases can expel the mobile phase from the pores of the stationary phase. This process is called dewetting and it adversely affects chromatographic performance. By design, the mildly hydrophilic surface of the Acclaim PA remains in contact with aqueous-only mobile phases, negating the problem of dewetting. While the onset of dewetting is somewhat unpredictable, stopping the flow of mobile phase through the column can initiate the process. The accompanying figure shows the effect of repeatedly stopping the flow through a C8 and a PA column. The C8 column dewets in a single cycle, but the PA remains wetted through many cycles. Protocol: Column: Acclaim PA, µm Dimensions:.6 mm Mobile Phase:. mm methanesulfonic acid, ph.6 Temperature: C Flow Rate: ml/min Injection Vol: µl Detection: UV, nm Each cycle consists of two steps:. Equilibrate column for min before testing for min.. Stop flow for min before next cycle begins.. Cytosine. Uracil. Thymine Physical Specifications Bonding C6 monomeric, polar-embedded USP code na Endcapping Double % C Pore size Å Surface area 9 m /g Particle diameters.9. µm or.. µm Hydrolytic Stability In order to reduce tailing and improve the peak shape of amine-containing compounds, HPLC separations of polar analytes are often run under acidic conditions. Separations under these conditions can shorten column life due to cleavage of the bonded phase. This cleavage results in frequent column replacement and instrument downtime. The proprietary bonding of the Acclaim PA column resists hydrolytic attack by protecting the bonded phase at low ph values

19 Acclaim PolarAdvantage Relative Retention (%) Hydrolytic Stability Comparison at Low ph Acclaim PolarAdvantage Brand C Brand B Columns: -µm polar-embedded Dimensions:.6 mm Mobile Phase: : v/v % TFA, ph.:ch CN Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: UV, nm Analyte: Toluene Performance Specifications Lot Polar selectivity ratio..8 Base asymmetry.98. ( µm).98.6 ( µm) Metal activity ratio.8.9 Acid asymmetry.98.6 Column Efficiency (N/m) 9 ( µm) ( µm) Asymmetry (EP).98. Retention time ( µm) ( µm) Time of Purge (days) 9 Example chromatographic specifications for Acclaim PA, C6,.6 mm. Please refer to Part for a detailed description of the tests and specifications. Specifications are subject to change at the discretion of Dionex. Chart of retention vs time at % acetonitrile,.% TFA for polar-embedded columns. LC/MS Compatibility Acclaim PA may be used for LC/MS applications. For best results and lowest background, operate below % organic solvent and between ph and ph 7. A new column should be conditioned for one hour with % acetonitrile. Store the column in % organic solvent between uses, and flush the column to waste with organic solvent before each use. rdering Information: Acclaim PolarAdvantage (PA) PA Standard particle sizes are nominally and µm. Analytical columns are stocked in.- and.6-mm-diameter widths and a range of lengths. Guard cartridges in both.- and.-mm sizes, packed with -µm particles, are recommended to protect both the and µm analytical columns. Capillary formats are available in 7 and µm,.-mm and custom diameters, and in -, -, or -mm lengths. Micro precolumn and nano precolumn cartridges are available in several formats. ther geometries can be made to order. Inquire with your Dionex sales representative. Please order through your local Dionex office or distributor. Refer to the following part numbers. Acclaim PA Analytical Columns µm. mm µm. mm µm. mm µm.6 mm µm.6 mm µm.6 mm... 6 µm.6 mm... 6 Acclaim PA Guard Columns µm. mm....6 µm. mm....6 Biocompatible cartridge holder for. and. mm guards...96 Guard to analytical column coupler...97 Holder and coupler kit Acclaim PA Micro and Nano Precolumns µm µm mm (pack of )...6 µm µm mm (pack of )...6 µm µm mm (pack of )...66 µm µm mm (pack of )...67 µm 8 µm mm (pack of )...6 µm mm (pack of )....6 µm mm (pack of ) Acclaim PA Micro and Nano Columns µm 7 µm mm (NAN7)... 6 µm 7 µm mm (NAN7) µm µm mm (FUS) µm µm mm (FUS) µm mm (MIC) µm mm (MIC) µm 7 µm mm (NAN7)... 6 µm 7 µm mm (NAN7) µm 7 µm mm (NAN7) µm µm mm (FUS) µm µm mm (FUS) µm µm mm (FUS) µm mm (MIC) µm mm (MIC) µm mm (MIC) These columns are designed for optimal performance using Dionex UltiMate and ICS- chromatography instruments

20 Acclaim PolarAdvantage II PA Chapter : Acclaim PolarAdvantage II (PA) Polar-embedded reversed-phase columns with enhanced hydrolytic stability Acclaim PA The Acclaim PA, like the Acclaim PA, is a high-efficiency, silica-based, reversed phase column with a polar enhanced stationary phase for operation over a wider range of chromatographic conditions than is possible with conventional reversed phase stationary phases. The Acclaim PA is an amide polar-embedded phase, with all the advantages of conventional polar-embedded phases, but with enhanced hydrolytic stability at both low and high ph (ph.-). The Acclaim PA provides selectivity that is complementary to conventional C8 columns, and to our Acclaim PA, providing a well-rounded column portfolio for methods development. This column is fully compatible with % aqueous mobile phases and provides symmetrical peaks for both polar and non-polar analytes. Si Unique Selectivity of the Acclaim PA (CH ) Si (CH ) Acclaim C8 Acclaim PA 6 N C (CH ) 6 CH Column: See chromatograms, µm Dimensions:.6 mm Mobile Phase: CHCN/ mm potassium phosphate buffer, ph. v/v 6/ Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: UV, 6 nm. Uracil 7. µg/ml. Pyridine. Phenol. N,N-Dimethylaniline. Toluene 6 6. p-butylbenzoic acid Polarity = [ K BBA K phenol /K toluene] 8 6 Acclaim C8 Surface chemistry USP code Endcapping Luna() C8 Acclaim PA SymmetryShield C8 8 Discovery RP-Amide C6 Physical Specifications ProntoSIL EPS C8 C8 polar-embedded na Yes %C. 7.% Pore size Å Surface area 6 m /g Particle diameters.9. µm or.. µm Recommended Ranges of peration 9 Acclaim PA The type 6 stainless steel column hardware is compatible with common HPLC mobile phases. To prevent corrosion, high concentrations of halide ions at ph <. should be avoided. ph Range. Pressure limit psi rganic solvents % 8 ZRBAX Bonus-RP The polarity index chart above ranks commercially available stationary phases according to their polarity relative to one another, under the same conditions. The more similar the polarity index of two stationary phases, the more similar their separation of polar compounds should be. If one column does not provide adequate resolution, choosing a column with a different polarity index may deliver a better separation. The Acclaim PA benefits include: Ability to separate polar and non-polar compounds Exceptional hydrolytic stability (ph. ) High polarity for complementary selectivity to C8 columns Compatibility with % aqueous or % organic solvent mobile phases Good peak shapes for both acidic and basic compounds High column efficiency Broad range of applications in pharmaceutical, environmental, food testing and product-quality testing. Resistance to Dewetting The carefully-designed bonding technology of the Acclaim PA incorporates hydrophilic functional groups between the hydrophobic alkyl chain and the silica surface that allows the surface to remain wetted, even in % aqueous mobile phase conditions. The accompanying figure shows the effect of repeatedly stopping the flow through a C8 and PA column. The C8 column is dewetted in a single cycle, but the PA provides reliable retention times for even very hydrophilic nucleic acid bases such as cytosine, uracil, and thymine, through many cycles. - -

21 Acclaim PolarAdvantage II Dewetting Test in % Aqueous Eluent Columns: µm,.6 x mm Mobile Phase:. mm methanesulfonic acid Temperature: C Flow Rate:. ml/min Conventional C8 Before stop-flow test After stop-flow test Inj. Volume: µl Detection: UV, nm. Cytosine. Uracil. Thymine Acclaim PA Before stop-flow test After stop-flow test % Remaining Retention Accelerated Stability Test at Low ph Competitor C Competitor A Competitor B Acclaim PA Time of Purge (Days) Column: See chart, µm Dimensions:.6 mm Mobile Phase: CH CN/% TFA*, ph v/v / Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: UV, nm Analyte: *Trifluoroacetic acid Toluene 6 8 Hydrolytic Stability 6 8 Despite their many advantages, the hydrolytic stability of polar embedded phases is often inferior to conventional C8 phases as a result of lower ligand coverage and the hydrophilic nature of the embedded group. The Acclaim PA has been designed to provide all the advantages of polar embedded phases, but with enhanced stability at both low and high ph. HPLC separations of polar analytes are often run at low ph (PH <.) to reduce tailing of amine-containing compounds. The accompanying figure shows that even at ph., the Acclaim PA is stable to hydrolysis unlike several other polar-embedded phases. Similarly, at ph. the Acclaim PA retains 9% of its retention capacity after 6 hours, while other columns with extended ph compatibility drop to -8%. Thus, the proprietary bonding of the Acclaim PA provides protection against hydrolytic attack at both low and high ph. 6 Performance Specifications Lot Polar selectivity ratio.8. Base asymmetry.98.6 Metal activity ratio.9.7 Acid asymmetry.98. Column Efficiency (N/m), ( µm) 6, ( µm) Asymmetry (EP).98. Retention time ( µm) ( µm) Example chromatographic specifications for Acclaim PA, C6,.6 mm. Please refer to Part for a detailed description of the tests and specifications. Specifications are subject to change at the discretion of Dionex. PA rdering Information: Acclaim PolarAdvantage II (PA) The standard particle sizes for the Acclaim PA columns are nominally - and -µm. Analytical columns are stocked in. and.6 µm diameter widths and a range of lengths. Guard cartridges in both.- and.-mm sizes, packed with -µm particles are recommended to protect both the - and -mm analytical columns. ther geometries can be made to order. Inquire with your Dionex sales representative. Please order through your local Dionex office or distributor. Refer to the following part numbers. Acclaim PA Analytical Columns -µm.6 mm µm.6 mm µm. mm µm.6 mm µm.6 mm Acclaim PA Guard Columns µm. mm (pack of ) µm. mm (pack of ) Biocompatible cartridge holder for. and. mm guards...96 Guard to analytical column coupler...97 Holder and coupler kit These columns are designed for optimal performance using Dionex UltiMate and ICS- chromatography instruments. - -

22 Acclaim A Chapter 6: Acclaim A for rganic Acids ptimized and application-tested for the analysis of hydrophilic organic acids Acclaim A organic acid columns are special reversed-phase silica columns, designed for the separation of aliphatic and aromatic organic acids at low ph with UV-Visible detection. The Acclaim A uses a patented polar-embedded stationary phase that allows a broad range of operating conditions, including % aqueous mobile phases. The Acclaim A is the recommended column for determining small hydrophilic organic acids, C to C7 aliphatic acids, and hydrophilic aromatic acid and is also valuable for the analysis and quality assurance of food and beverage products, pharmaceutical preparations, plating baths for semiconductor manufacturing, manufacturing chemicals, and chemical intermediates. The Acclaim A columns are packed in metal-free PEEK column bodies, to eliminate unwanted interactions between the analytes and the column body. Benefits of the Acclaim A columns include: Use-tested to guarantee consistent organic acid separations Compatible with % aqueous mobile phases, optimum for reversed-phase retention of hydrophilic aliphatic organic acids Hydrolytic stability at low-ph conditions, optimum for reversed-phase retention of organic acids Ideal selectivity for separating a wide spectrum of organic acids, including hydrophilic aliphatic acids, C to C7 aliphatic acids, aromatic acids, and selected amino acids Excellent peak shapes for organic acids High reversed-phase capacity with retention similar to standard C8 phases Suitable for analysis of foods, beverages, pharmaceuticals, chemical intermediates, and environmental samples Ideal for quality assurance offering fast run times and minimal sample preparation A Recommended Ranges of peration ph Range. 8. Pressure limit psi ( psi above % organic solvent content) rganic solvents Hydrophilic rganic Acids H H H H H H H H H H H H H H. xalic acid. Tartaric acid. Formc acid. Malic acid. iso-citric acid H H H H H H H H H C H C H H H 6. Lactic acid 7. Acetic acid 8. Citric acid 9. Succinic acid. Fumaric acid H H H H H. cis-acontic acid. trans-aconitic acid H % acetonitrile or methanol 9 Surface chemistry USP code Endcapping Pore size Physical Specifications Proprietary na Yes Å Surface area 9 m /g Particle diameter.. µm Guaranteed Performance for rganic Acids The performance of the Acclaim A columns is guaranteed for the separation of aliphatic and aromatic organic acids at low ph. These columns undergo extensive testing to ensure columnto-column reproducibility, and are shipped with quality assurance reports detailing these tests. The Acclaim A columns are usetested for two specific applications isocratic and gradient. These operating conditions provide a good starting point for methods development. Performance Specifications Lot Metal activity ratio.8. Gradient organic acids chromatogram Column Efficiency (N/m) 7, Asymmetry.9.7 Retention time Resolution (Lactic-Acetic).8 Baseline drift < m Resolution (Caproic-isocaproic).8 Example chromatographic specifications for Acclaim A. mm. Please refer to Part for a detailed description of the tests and specifications. Specifications are subject to change at the discretion of Dionex. - -

23 Acclaim A Isocratic Separation of Hydrophilic rganic Acids Ruggedness Test at ph Column: Acclaim A, µm Dimensions: mm Mobile Phase: mm Na S, ph.6 (adjusted with methanesulfonic acid) Temperature: C Flow Rate:.6 ml/min Inj. Volume: µl Detection: UV, nm. xalic acid mg/l (ppm). Tartaric acid. Formic acid 8. Malic acid. iso-citric acid 6. Lactic acid 8 7. Acetic acid 8. Citric acid 9. Succinic acid. Fumaric acid 7. cis-aconitic acid **. trans-aconitic acid ** ** 7 ppm total for cis and trans isomers Injection Injection Column: Acclaim A, µm Dimensions: mm Mobile Phase: mm Na S, ph (adjusted with methanesulfonic acid) Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: UV, nm. xalic acid mg/l (ppm). Tartaric acid. Formic acid 8. Malic acid. iso-citric acid 6. Lactic acid 8 7. Acetic acid 8. Citric acid 9. Succinic acid. Fumaric acid. cis-aconitic acid**. trans-aconitic acid** * * ppm total for cis and trans isomers Gradient Separation of C C7 Aliphatic rganic Acids.8., Column: Acclaim A, µm Dimensions: mm Mobile Phases: A: CH CN B:. mm methanesulfonic acid Gradient: - min. 6 6 %A %B Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: UV, nm with blank subtraction. Formic acid mmol/l. Acetic acid. Propionic acid. Butyric acid. Isobutyric acid 6. Isovaleric acid 7. n-valeric acid 8. Isocaproic acid 9. n-caproic acid. Heptanoic acid A 6 rdering Information: Acclaim A The standard particle size is nominally µm. Analytical columns are available in.-mm diameters; standard lengths are and mm. Guard cartridges in. mm are recommended to protect your analytical columns. ther geometries in PEEK or stainless steel are available; inquire with your Dionex sales representative. Please order through your local Dionex office or distributor. Refer to the following part numbers. Acclaim A Analytical Columns µm. mm µm. mm Acclaim A Guard Columns µm. mm Biocompatible cartridge holder for. mm and. mm guards...96 Guard to analytical column coupler...97 Holder and coupler kit These columns are designed for optimal performance using Dionex UltiMate and ICS- chromatography instruments. - -

24 Acclaim Surfactant Chapter 7: Acclaim Surfactant ptimized and application-tested for the analysis of anionic, nonionic and cationic surfactants The Acclaim Surfactant Column The Acclaim Surfactant is a silica-based, reversed phase column that features a proprietary bonding chemistry optimized and application-tested for separating a variety of surfactants. This specialty column is ideal for separating cationic, nonionic, anionic and amphoteric surfactants in a single chromatographic run using simple and volatile mobile phases. The Acclaim Surfactant column is the ideal tool for the analysis of surfactants including linear alkylsulfonates, alkyl sulfates, alkyl quaternary ammonium salts, ethoxylated quats, Triton-X, PEGs and many more. It allows analysis of surfactants in various matrices including consumer products, formulations for pharmaceutical products, and environmental samples. mv Inorganic Anions, Hydrotropes, Cationic, Nonionic, Amphoteric, and Anionic Surfactants Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) CH CN, (B). M NH Ac, ph. Gradient: % to 8% A in min, then hold 8% A for min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELS detector Chloride. Bromide. Nitrate. Xylene sulfonate. Laurylpyridinium chloride 6. Lauryldimethylbenzylammonium chloride 7. Triton X- 8. Cetyl betaine 9. Decyl sulfate. Dodecyl sulfate. C -LAS. C -LAS. C -LAS. C -LAS 79 Benefits of the Acclaim Surfactant column include: Ideal selectivity for separating different types of surfactants Excellent peak shapes, especially for cationic surfactants Improved resolution for ethoxylated surfactants compared with other columns Ability to analyze hydrophilic hydrotropes Methods compatible with various detection methods including UV, ELSD, suppressed conductivity, and more Broad range of applications Hydrotrope, Cationic, Nonionic, Amphoteric and Anionic Surfactants mv µs Separation of Commonly Used Cationic Surfactants ELS Detector Suppressed Conductivity Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) CH CN (B) mm Acetic acid Gradient: % A in min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELS detector or Suppressed CD (CMMS III). Lauryl pyridinium chloride. Lauryldimethylbenzyl ammonium chloride. ctylphenoxyethoxyethyl dimethylbenzyl ammonium chloride. Cetyltrimethylammonium chloride. Cetylpyridinium chloride SURFACTANT CH CH S Na Sodium Xylene Sulfonate + N Cl Laurylpyridinium Chloride 86 CH CH N + CH Cl Lauryldimethylbenzyl Ammonium Chloride Cetyl Betaine Sodium Dodecyl Sulfate -S-Na Me + N Me C 8 H 7 Triton X- H (CH CH ) n H n ~ Sodium Decyl Sulfate CH (CH ) n C (CH ) m CH S Na m + n = Sodium Dodecylbenzene Sulfonates -S-Na 79 Bonding USP code Pore size Physical Specifications Proprietary na Å Surface area m /g Particle diameters.. µm Recommended Ranges of peration The type 6 stainless steel column hardware is compatible with common HPLC mobile phases. To prevent corrosion, high concentrations of halide ions at ph <. should be avoided. ph Range. 7. Pressure limit psi rganic solvents % - -

25 Acclaim Surfactant Guaranteed Performance for Surfactants The exceptional performance of Acclaim Surfactant columns for the separation of anionic, cationic and non-ionic surfactants is confirmed with every new lot of bonded silica that is manufactured. In addition each column is individually tested with a simple mixture of anionic, cationic and non-ionic probes, to ensure that every column is packed consistently and well. Performance Specifications Column Efficiency (N/m) 76, Asymmetry (EP).9. Retention time Example chromatographic specifications for Acclaim Surfactant.6 mm. Please refer to Part for a detailed description of the tests and specifications. Specifications are subject to change at the discretion of Dionex. 9 LT VALIDATIN #6 #, SFC_P (isocratic) V little or no modifications, because both detectors share the same mobile phase requirements. Conductivity detection of cationic surfactants can also be used. Because of the versatility of this column, methods using all these detectors have been developed and can be found in this catalog, in the Acclaim product information bulletin, or on the Dionex website. Abundance Analysis of PE () Lauryl Sulfate (LC-ESI-MS) :7 :8 :9 : :6 : : : : : (alkyl chain: n) : Column: Acclaim Surfactant, µm Dimension:.6 mm Mobile Phase: CH CN/. M NH Ac, ph. v/v 6/ Temperature: C Flow Rate: ml/min (. ml/min to MS) Inj. Volume: µl Detection: MS (ESI negative) Sample: PE () lauryl sulfate ( ppm) : : n ~ Sodium PE () Lauryl Sulfate (CH CH ) n - -S- Na 79 - min.... Chromatogram of a group of surfactants, taken from the certificate for lot #--. Dionex monitors and controls the selectivity of cationic, nonionic and anionic surfactants using the recommended standard conditions for this column. Detector Compatibility Although UV detection is the most commonly used of HPLC detectors, not all surfactants possess chromophores. This problem can be overcome by the use of a universal detector, such as evaporative light-scattering detection (ELSD) or refractive index (RI) detection. ELSD is compatible with gradient methods and is far more sensitive than RI. In addition, methods developed with ELSD can be easily transferred to LC-ESI-MS applications with Dimensions:.6 mm Mobile Phases: Acetonitrile/ mm NHAc, ph., v/v 7/ for Acclaim Surfactant Acclaim Surfactant C-LAS v/v / for Acclaim PA C-LAS Temperature: C C-LAS Flow Rate: ml/min Inj. Volume: µl C-LAS Detection: UV, nm C-LAS C-LAS C-LAS Acclaim PA H C-LAS CH (CH ) n C (CH ) m CH Analyis of Sodium Dodecylbenzene Sulfonate (LAS) S Na m + n =7- Sodium Dodecylbenzene Sulfonate 78 SURFACTANT rdering Information: Acclaim Surfactant Standard particle size is nominally -mm. Analytical columns are stocked in.6 mm-diameter widths and a couple of lengths. Guard cartridges in.-mm sizes, packed with -mm particles are recommended to protect the analytical columns. ther geometries can be made to order. Enquire with your Dionex sales representative. Please order through your local Dionex office or distributor. Refer to the following part numbers. Acclaim Surfactant Analytical Columns µm.6 mm... 6 µm.6 mm... 6 Acclaim Surfactant Guard Columns µm. mm (pack of )....6 Biocompatible guard cartridge holder...96 Guard for analytical column coupler Acclaim holder and coupler kit...96 These columns are designed for optimal performance using Dionex UltiMate and ICS- chromatography instruments. - -

26 Acclaim Explosives Chapter 8: Acclaim Explosives E and E Columns For the separation of the explosives targeted by EPA SW-86 Method 8 The novel and unique chemistries of these columns provide superior resolution to other commercially available columns. The Acclaim Explosives columns provide a "Total Solution" to EPA Method 8; two columns each of which provides baseline resolution of all explosives. The Acclaim Explosives E provides selectivity similar to conventional C8 columns. The Acclaim E column provides selectivity that is complementary to that obtained using the Acclaim E and is recommended for the confirmatory analysis. With their complementary selectivities, the E and E provide both primary and confirmatory columns. Acclaim Explosives Columns The Acclaim Explosives E and E are specialty reversed phase columns that have been optimized and application tested for the separation of the explosives targeted by EPA SW-86 Method 8, Nitroaromatics and Ntramines by HPLC. Baseline Separation of Target Explosive Compounds Listed in EPA SW-86 Method 8. HMX. RDX.,,-Trinitrobenzene.,-Dinitrobenzene. Nitrobenzene 6. Tetryl 7.,,6-Trinitrotoluene 8. -Amino-,6-Dinitrotoluene 9. -Amino-,6-Dinitrotoluene.,6-Dinitrotoluene.,-Dinitrotoluene. -Nitrotoluene. -Nitrotoluene. -Nitrotoluene Fourteen Target Explosives Listed in EPA Method 8 CH N N H C N N N N N N N N N N N N N N N N N N N N TNT HMX RDX Tetryl CH CH N N N N N Acclaim Explosives E Acclaim Explosives E 8 9 N N N,6-DNT,-DNT,-DNB NB CH CH N NH N N N N Benefits of the Acclaim Explosives E and E columns N -A-,6-DNT NH -A-,6-DNT,,-TNB Superior resolution for all explosives listed in EPA Method 8 CH N CH CH Complementary selectivity between Acclaim E and E, for a complete solution for explosives analysis EXPLSIVES N -NT -NT -NT N 77 -µm, Å high purity silica gel for high-resolution separations, with good mechanical stability Unique selectivity of the Acclaim E allows for simultaneous separation of some nitrate ester explosives - 6 -

27 Acclaim Explosives Separation of 8 Explosives Showing the Unique Selectivity of the Acclaim Explosives E Column Column: Acclaim Explosives E, µm Dimensions:.6 mm Mobile Phase: 8/ v/v MeH/H Temperature: ºC Flow Rate: ml/min Inj. Volume: ml Detection: UV, nm Physical Specifications Acclaim Explosives E Bonding Proprietary Proprietary Pore size Å Å Surface area m /g m /g Particle diameters..-µm..µm ( ppm each).,-di-amino-6-nitrotoluene. HMX. EGDN. RDX.,,-Trinitrobenzene 6.,-Dinitrobenzene 7. Nitrobenzene 8.,,6-Trinitrotoluene 9. Tetryl.,6-Dinitrotoluene.,-Dinitrotoluene. -Nitrotoluene. -Nitrotoluene. -Nitrotoluene. -Amino-,6-Dinitrotoluene 6. -Amino-,6-Dinitrotoluene 7. DMDNB 8. PETN 6 Acclaim Explosives E Recommended Ranges of peration These operating conditions follow EPA Method Conditions. Acclaim Explosives E Acclaim Explosives E Mobile phase MeH:H :7 MeH:H 8: Temperature ± C 9 ± C Flow rate. ml/min. ml/min Guaranteed Performance for EPA Method 8 The exceptional performance of both Acclaim Explosives columns for EPA Method 8 is guaranteed. Both products undergo extensive testing to ensure column-to-column reproducibility and are use-tested for this specific application. Both columns perform reproducibly under their optimized conditions, although separation conditions for the Acclaim Explosives E can be readily changed to accommodate inclusion of additional explosives compounds. Performance Specifications Acclaim Explosives E Efficiency (N),, Asymmetry Retention time Acclaim Explosives E Example chromatographic specifications for Acclaim E and E.6 mm. Please refer to Part for a detailed description of the tests and specifications. Specifications are subject to change at the discretion of Dionex rdering Information: Acclaim Explosives Columns Standard particle size is nominally -µm. analytical columns are stocked in.6 mm-diameter widths and -mm length. Guard cartridges in.-mm sizes, packed with -µm particles are recommended to protect the analytical columns. ther geometries can be made to order. Enquire with your Dionex sales representative. Please order through your local Dionex office or distributor. Refer to the following part numbers. Acclaim Explosives Analytical Columns Acclaim Explosives Guard Columns µm.6 mm E... 6 µm. mm E (pack of )....6 µm.6 mm E µm. mm E (pack of ) µm E and E kit... 6 Biocompatible guard cartridge holder...96 Guard for analytical column coupler Acclaim holder and coupler kit...96 These columns are designed for optimal performance using Dionex UltiMate and ICS- chromatography instruments. EXPLSIVES - 7 -

28 Applications Part Three: Applications Contents APPLICATINS Pharmaceutical Isocratic Resolution of Antihistamines and Their Impurities on Acclaim Å, C Separation of Four Benzodiazepine Drugs on Acclaim C Separation of Five Antidepressant on Acclaim PA... 9 Spectinomycin and Lincomycin on Acclaim PA with Electrochemical Detection... 9 Beta Blockers on Acclaim PA... 9 Venlafaxine and Related Substances... 9 Assay for Pramoxine in Topical Anesthetic intment on Acclaim Å, C8 with SPE Cleanup on nguard II RP... Determination of Four Active Ingredients in a Multisymptom Cold Remedy Using Acclaim PA. Separation of Basic Drugs on Acclaim Å, C8... Baseline Separation of Catecholamines & Serotonin at Physiological Concentrations on Acclaim Å, C8 with Electrochemical Detection... Gradient Resolution of Sulfa Drugs on Acclaim PA... Catecholoamines in Urine on Acclaim PA... Bromide and rganic Acids in Cough Syrup Using Acclaim A... Isocratic Separation of Six Cardiac Anti- Arrhythmic Drugs (Beta-Blockers) on Å C8... Sulfa Drugs on Acclaim PA... Active Ingredients in Zantac 7 Acid-Reducer Medication... Loratidine and Pseudoephedrine on Acclaim PA. Propafenone and Related Substances... Antidepressants on Acclaim PA... Active ingredients in Cough Syrup... Barbiturates on Acclaim C8... Budesonide and Related Substances... Hydrocortisone in Skin intment... Phenytoin and Related Substances... Active Ingredients in Sunscreen Lotion on Acclaim Å, C8 Using ASE Extraction... Glucocorticosteroids in Serum on Acclaim C8. Nonsteroidal Anti-inflammatory Drugs... Fexofenadine & Pseudoephedrine on Acclaim PA... Environmental LC/MS Detection of Herbicides on Acclaim C8... LC Separation of Nitrosamines in EPA 87 Using Acclaim PA... Benzidines by EPA Method 6 on Acclaim C8 with Electrochemical Detection... Phenol Standards by EPA Method 6 on Acclaim PA... Separation of Phthalate Esters in EPA Method 66 Using Acclaim PA... Brodifacoum in Rodent Bait... Thirteen Carbonyl-DNPH Derivatives for CARB Method on Acclaim Å, C8... Carbamate Insecticides in EPA. on Acclaim C8 with MS Detection... Improved Separation of Carbonyl-DNPH Derivatives by CARB Method Using Coupled Acclaim PA and Acclaim C8 Columns... Polynuclear Aromatic Hydrocarbon Standards by EPA Method 8 on Acclaim PA... LC Separation of Benzidines in EPA 87 Using Acclaim PA... 6 Sensitive Detection of Triazine Herbicides on Acclaim C8 with MS Detection... 6 EPA Method 8 Using Acclaim Explosives E.. 6 EPA Method 8 Using Acclaim Explosives E.. 6 Four Antibiotics in Animal Feed on Acclaim C Carbamate Insecticide Standards in EPA. on Acclaim PA... 6 Food and Beverage Fat-Soluble Vitamin Standards on Acclaim C Amino Acids in Vitamin Premix on Acclaim A.. 7 rganic Acids in range Juice on Acclaim A... 7 rganic Acids in White Wine on Acclaim A... 7 Carotenoids in Serum on Acclaim PA... 7 Pigments in Turmeric on Acclaim PA... 7 Assay of Aspartame and Its Impurities on Three Formats of Acclaim C8 Columns... 8 Assay for Water-Soluble Vitamins in Vitamin Tablets Using Acclaim PA... 8 Nitrofuran Antibiotic Residues in Animal Feed... 8 Fat-Soluble Vitamins and Carotenoids in Vitamin Tablets... 8 Determination of Quinine in Tonic Water... 9 Determination of Ingredients of an Energy Drink. 9 Hops Extract on Acclaim C Sudan Dyes in Paprika... 9 Water-Soluble Vitamins Using Acclaim PA... 9 Parabens on Acclaim PA... 9 Four Aflatoxin Standards on Acclaim C8... Fat-Soluble Vitamin Standards Using Acclaim PA... Glutamate in Beer by Automated Precolumn PA Derivatization... Determination of Soft Drink Ingredients... Tetracycline Antibiotic Residues in Pork... Fat-Soluble Vitamins and Carotenoids in Dry Pet Food... Antioxidants in Edible ils by AAC 98. on Acclaim C8... Sulfonamide Anibiotic Residues in Milk... Bitter Principles in Beer on Acclaim C8... Chemical Separation of Cationic Surfactants Using ELSD... Separation of Ethoxylated Quats... Analysis of ZNYL FS Fluorosurfactant... Analysis of a Shampoo... Analysis of a Fabric Softener... Analysis of a Mouthwash... Separation Hydrotropes, Anionic, Cations, and Amphoteric Surfactiants... Analysis of Sodium Lauryl Sulfate Using LC-ESI-MS... Analysis of PEG monoethyl ether (MW-)... Analysis of Triton X- Using LC-ESI-MS... Analysis of a Laundry Washing Detergent... Analysis of a Dishwashing Liquid... Analysis of Ammonium Laureth Sulfate Using Conductivity Detection... Separation of Inorganic Anion, Hydrotropes, Cations, Nonionic, Amphoteric and Anionic Surfactiants... Analusis of Toilet Bowl Cleaner... Analysis of a Strongly Hydrophilic Hydrotrope... Analysis of Quats Using LC-ESI-MS... Analysis of NEDL Analysis of PEG- Distearate... Separation of Cationic Surfactants with Suppressed Conductivity Detection... Separation of Quaternary Imidazolinium Compounds... Separation of Different Polyethylene Glycols... PE() Ammonium Lauryl Sulfate... Sudan Dyes on Acclaim C8... Bioscience Tryptic Peptide Map of Cytochrome c on Acclaim C Capillary LC for Peptide Mapping Using Acclaim C8...6 Separation of Protein Standards by Micro-LC on Acclaim C Tryptic Peptide Map of Bovine Serum Albumin on Acclaim C Cytochrome c from Three Species on Acclaim C Protein Mixture by Reversed-Phase HPLC on Acclaim C Peptide Test Mix on Acclaim and PA with Formic Acid... 7 Comparison of Acid Additives for Peptide Analysis... 7 Natural Products Alkaloids in Goldenseal Root on Acclaim C8 with ASE Extraction... 8 Isoflavones in Red Clover on Acclaim C8 with ASE Extraction... 8 Flavonoids in Green and Black Teas Using Acclaim C rganic Acids Resolution of Benzene Polycarboxylic Acids on Acclaim A... 9 Acrylic Acid ligomers on Acclaim A... 9 Separation of Hydroxybenzoic Acids on Acclaim A

29 Pharmaceutical Applications Pharmaceutical m Isocratic Resolution of Antihistamines and Their Impurities on Acclaim C8 6 8 Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: (A) mm sodium acetate (B) Methanol Isocratic: (A) %, (B) 8% Temperature: C Detection: UV, 9 nm. Thenyldiamine HCI. Phenothiazine. Promethazine HCI. Pyrrobutamine phosphate Beta Blockers on Acclaim PA 7 8 Column: Acclaim PA, µm Dimensions:.6 mm Mobile Phase: MeH/.% NH H, ph, v/v, 6/ Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: UV, nm ( ppm each). Maleate. Labetalol. Metaraminol. Atenolol. Acebutolol 6. Metoprolol 7. Timolol 8. xprenolol Separation of Four Benzodiazepine Drugs on Acclaim C8 m Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: 6% acetonitrile Flow Rate:. ml/min Detection: UV, nm. xazepam. Clonazepam. Temazepam. Diazepam 7 m Separation of Five Antidepressants on Acclaim PA Column: Acclaim PA, µm Dimensions:.6 mm Mobile Phase: 8/ v/v MeH/ mm phosphate, ph 6. Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: UV, nm. Protriptyline. Nortriptyline. Doxepin. Imipramine. Amitriptyline nc Spectinomycin and Lincomycin on Acclaim PA with Electrochemical Detection Column: Acclaim PA, µm Dimensions:. mm Mobile Phase (A) mm acetic acid,. g/l pentanesulfonic acid, ph. with NaH (B) mm acetic acid,. g/l pentanesulfonic acid, ph. with NaH (C) Acetonitrile Gradient: Time % A % B % C Flow Rate:. ml/min Temperature: C Inj. Volume: µl Detection: Postcolumn addition of. M NaH at. ml/min; pulsed amperometric detection. Benzyl alcohol. Spectinomycin anomer. Spectinomycin anomer. Spectinomycin. Lincomycin 8 m Venlafaxine and Related Substances Data courtesy of: V. Bhate of Analytical Solutions Column: Acclaim C8, µm,.6 mm Pump: Summit P68A Mobile Phase: (A) % acetonitrile; 9% water;. ml/l triethylamine, adj. to ph. with phosphoric acid Mobile Phase: (B) 7% acetonitrile; % water;. ml/l triethylamine, adj. to ph. with phosphoric acid Flow:. ml/min Gradient Times: 7 %A: %B: Temperature: TCC thermostat at C Injection: ASI- autosampler; µl Detector: UVD U; UV at 6 nm Sample: Resolution test mix. Impurity µg/ml. Impurity A. FL II. Venlafaxine. Impurity III 6. Impurity C 7. Impurity B 8. PMPA 9. Impurity IV. FL I. Impurity V APPLICATINS

30 Pharmaceutical Applications Separation of Basic Drugs on Acclaim C8 C B A Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: 8/ methanol/ mm phosphate, ph 6 Temperature : C Flow Rate: ml/min Inj. Volume: µl Detection: UV, nm Amitriptyline Mass: Trace A: ng Trace B: ng (normalized peak height) Trace C: 9 ng (normalized peak height). Uracil. Propranolol. Toluene. Doxepin. Amitriptyline Note: The high performance of this column is maintained as the sample is diluted m Gradient Resolution of Sulfa Drugs on Acclaim PA Column: Acclaim PA, µm Dimensions.6 mm Mobile Phase: (A) 9/ methanol/ mm phosphate, ph.7 (B) /9 methanol/ mm phosphate, ph.7 Inj. Volume: µl Gradient: (A) % at min, to 7% at min, to 8% at min Temperature: C Flow Rate: ml/min Detection: UV, nm. Sulfanilic acid 6 µg. Sulfanilamide. Sulfadiazine. Sulfathiazole. Sulfamerazine 6. Sulfamethazine 7. Sulfamethoxazole 8. Sulfisoxazole 9. Sulfadimethoxine na Baseline Separation of Catecholamines and Serotonin at Physiological Concentrations on Acclaim C8 with Electrochemical Detection Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: 7 mm citric acid, mm sodium acetate,. mm EDTA, mm sodium octanesulfonate, % methanol Flow Rate: ml/min Detection: dc amperometry, Glassy Carbon at 7 mv, Ag/AgCl Referemce electrode. Norepinephrine pmol. Epinephrine. Dihydroxybenzylamine. Dopamine. Serotonin 88 m Sample Prep:. Disperse. g ointment in 8. ml MeH. Add. ml conc. NH H and make to ml with MeH. Apply. ml to a conditioned nguard II RP -cc cartridge and expel with ml air. Elute twice with. ml of % acetic acid in acetonitrile and expel with ml air.. Combine and make to ml Assay for Pramoxine in Topical Anesthetic intment on Acclaim C8 with SPE Cleanup on nguard II RP Pramoxine 7 Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: (A) MeH (B) mm H P, adjusted to ph. with NH H Gradient: %A until.7 min, ramp to 8% A at 6 min, end at 7 min Flow Rate:. ml/min Temperature: C Detection: UV, 6 nm Inj. Volume: µl 96 APPLICATINS m Determination of Four Active Ingredients in a Multisymptom Cold Remedy Using Acclaim PA 7 Column: Acclaim PA, µm Dimensions:.6 mm Injection: µl Mobile Phase: (A) Acetonitrile (B) mm phosphoric acid adjusted to ph. with ammonium hydroxide Temperature: C Detection: UV, nm Gradient : Time % A % B Flow Peak # Compound Label Concentration % Recovery Pseudoephedrine HCl. mg/ml 96 Acetaminophen Doxylamine succinate. 9 Dextromethorphan HBr na Catecholamines in Urine on Acclaim PA Column: Acclaim PA, µm Dimensions:. mm Mobile Phase: Buffer (.98 g citric acid,. g NaAc, 7. mg EDTA, ml of. M methanesulfonic acid in L D.I. water)/meh v/v 9/ Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: DC amperometry [GC electrode, 8 mv] ( nm each). -Hydroxy--methoxymandelic acid. -Hydroxy--methoxyphenylglycol. Norepinephrine. Epinephrine.,-digydroxybenzlamine 6. Normetanephrine 7. Metanephrine 8. Dopamine 9. -Hydroxy--methoxyphenylacetic acid. -Hydroxyindoleacetic acid

31 Pharmaceutical Applications. Bromide and rganic Acids in Cough Syrup Using the Acclaim A Column: Acclaim A, µm Dimensions: mm Mobile Phase:. M Na S, ph.68 (adjusted with methanesulfonic acid) Temperature: C Flow Rate:.6 ml/min Inj. Volume: µl Detection: UV, nm Sample Prep: nguard II P Sample: NyQuil. Bromide (Dextromethorphan HBr). Citrate (Inactive ingredient). Succinate (Doxylamine succinate) NyQuil is a registered trademark of Proctor and Gamble Sulfa Drugs on Acclaim PA 8 9 Column: Acclaim PA, µm Dimensions:.6 mm Mobile Phase: CH CN/. M NH Ac, ph., v/v, /7 Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: UV, 6 nm ( ppm each). Sulfanilamide. Sulfathiazole. Sulfamerazine. Sulfamethazine. Sulfisoxazole 6. Sulfapyridazine 7. Sulfamethoxazole 8. Sulfadimethoxine 9. Sulfaquinoxaline sodium Loratidine and Pseudo-ephedrine in a Time-Release Tablet on Acclaim PA Column: Acclaim PA C6, µm,.6 mm Pump: Summit P68A DGP-6 Mobile Phases: (A) :8:. acetonitrile:water:tfa (B) :6:. acetonitrile:water:tfa m Flow:. ml/min Temperature: C Gradient: Times nm %A %B Injection: Summit ASI-, µl Detector: Summit UVD U diode array, UV at 6, and 7 nm Sample Preparation: Finely grind a weighed tablet. Weigh an aliquot containing mg loratidine ( mg) into a dry ml flask. Add ml methanol and disperse. Add ml of N HCl with mixing. Dilute to ml with water. Filter through. µm membrane filter.. Pseudo-ephedrine µg/ml. Loratidine 78 6 m B A Propafenone and Related Substances 6 Column: Acclaim PA C6, µm,.6 mm Pump: Summit P68A LPG Mobile Phase: (A). M ammonium acetate, adj. to ph. with phosphoric acid (B) Methanol Flow:. ml/min Gradient Times: %A: 8 8 %B: Temperature: TCC thermostat at C Injection: ASI- autosampler; µl Detector: UVD U; UV at 9 nm Sample Preparation: Weigh accurately about mg of sample in ml standard volumetric flask. Add 6 ml of methanol and sonicate for min and dilute up to the mark with methanol. Centrifuge the above solution for min and further dilute ml of clear supernatant solution to ml with diluent. Diluent is mobile phases A:B :6. Data courtesy of: V. Bhate of Analytical Solutions Sample: (A) Resolution test mix (B) Formulation. Propafenone µg/ml. Epoxyefenone. Chlorohydrine 8. Tertiary amine. HBAH 6. Dimer 66. Isocratic Separation of Six Cardiac Antiarrhythmic Drugs (Beta-Blockers) on Acclaim C Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: /9 w/w MeH/ mm phosphate, ph 7. Flow Rate:. ml/min Temperature: C Inj. Volume: µl Detection: UV, nm. Maleic acid. Acebutolol µg/ml. Metoprolol. Timolol. xprenolol 6, 7. Labetalol diasteromers 8. Propranolol Active Ingredient in Zantac 7 Acid-Reducer Medication Reference: Ho C, Huang HM, Hsu SY, Shaw CY, Chang BL, Drug Dev. Ind. Pharm. 999 :79-8. Zantac is a registered trademark of Warner-Lambert Co. Column: Acclaim C8, µm,.6 mm Pump: GP micro Mobile Phase: mm KH P : acetonitrile:methanol: triethylamine :::.7 v:v:v:v isocratic Flow:. ml/min Temperature: C Inj. Volume: µl Detector: AD UV/VIS at nm Sample Prep.: ne tablet dissolved in ml water; filtered. Ranitidine. Ranitidine S-oxide (impurity) APPLICATINS - -

32 Pharmaceutical Applications m FS 6 9 m FS Budesonide and Related Substances Reference: Hou S, Hindle M, Byron PR; J. Pharm. Biomed. Analy. : Column: Acclaim C8, µm,.6 x mm Pump: Summit P8 HPG/ Mobile Phase: (A) Acetonitrile:ethanol : (B).% phosphoric acid Isocratic 66% B Flow:. ml/min Temperature: C Injection: ASI- autosampler, µl Detector: UVD U; UV at nm Sample: Budesonide, µg/ml after three days 7, 8. Budesonide epimers, 99%.. Hydrocortisone in Skin intment 6 Column: Acclaim C8, µm,.6 mm Pump: Dionex GP microbore Mobile Phase: Acetonitrile:water 9: Flow:. ml/min Temperature: C Injection: µl Detector: AD-; UV at nm Sample Prep.: Disperse mg of ointment in 7. ml denatured ethanol Mix in. ml hexane then. ml water Centrifuge to separate emulsion Filter ethanol layer. Hydrocortisone.% (label.%). Methyl paraben.%. Propyl paraben.% 6 6 m Barbiturates on Acclaim C Column: Acclaim C8 µm,.6 x mm Pump: Summit P8 HPG/ Mobile Phase: (A) Acetonitrile:methanol : v:v (B).% phosphoric acid Gradient: Time.. 7. %A %B 8 Flow:. ml/min Temperature: C Injection: ASI- autosampler, µl Detector: UVD U, UV at nm Sample: µg/ml each in methanol. Barbital. Phenobarbital. Butabarbital. Amobarbital. Secobarbital.8 Active Ingredients in Cough Syrup Column: Acclaim PA, µm Dimensions:.6 mm Mobile Phase: Acetonitrile/ mm phosphate buffer, ph.9 from /9 to /6 (v/v) in min Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: UV, nm. Doxylamine. Pseudo-ephedrine. Acetaminophen. Dextromethorphan 6. Antidepressants on Acclaim PA Column: Acclaim PA, µm Dimensions:.6 mm Mobile Phase: MeH/ mm triethylamine, ph.8, v/v 7/ Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: UV, nm Concentration Asymmetry (µg/ml) (EP). Uracil.8. Doxepin.. Imipramine 8.. Amitriptyline. Phenytoin and Related Substances Column: Acclaim, C8 µm,.6 mm Pump: Summit P68A Mobile Phase: % methanol; % acetonitrile; m %. M NH H P adj. to ph. Flow:. ml/min B 6 Temperature: TCC thermostat at 6 C 7 8 Injection: ASI- autosampler; µl A Detector: UVD U; UV at nm Samples: (A) Low load test mix (B) Resolution test mix Data courtesy of: V. Bhate of Analytical Solutions. Impurity A µg/ml. Impurity B. Benzoic acid 9. Benzaldehyde 9. Phenytoin sodium (B) or (A) 6. Benzoin 9 7. Benzophenone 8. Benzil APPLICATINS - -

33 Pharmaceutical Applications Glucocorticosteroids in Serum on Acclaim C8 Nonsteroidal Anti-inflamatory Drugs on Acclaim PA C8 m B A 6 7 Reference: McWhinney B C, Ward G, Hickman P E; Clin. Chem, 996, : Column: Acclaim C8 µm,.6 mm Pump: Summit P8 HPG/ Mobile Phase: Methanol:tetrahydrofuran:water ::7 v:v:v, isocratic Flow:. ml/min Temperature: C Injection: ASI- autosampler, 6 µl Detector: UVD U; UV at nm Samples: (A) Extract of Bovine Serum extracted with ethyl acetate from serum alkalized with NaH (B) Six steroid standards, µg/ml in mobile phase. Prednisone. Prednisolone. Cortisol. Fludrocortisone (I.S.). Methylprednisolone 6. -Deoxycortisol 7. Dexamethasone Column: Acclaim PA C8, µm,.6 mm Pump: Summit P 68A DGP-6 Mobile Phase: Acetonitrile:.% phosphoric acid, : v/v; isocratic 9 m.. nm 98 Flow:. ml/min Temperature: C Injection: Summit ASI-, µl Detector: Summit UVDU diode array, 6nm, and spectra nm nm Ketoprofen µg/ml. Naproxen. Ibuprofen 6. nm 98,8 m Active Ingredients in Sunscreen Lotion on Acclaim C8 Using ASE Extraction Extraction Conditions: ASE with ml extraction cell / MeH/CH Cl Column: Acclaim C8 Mobile Phase: Isocratic 8//.7 methanol/water/hac Inj. Volume: µl Detection: UV, nm. Benzophenone-. ctyl methoxycinnamate. ctyl salicylate 98 m Fexofenadine and Pseudoephedrine on Acclaim PA * UV 8 nm * Column: Acclaim PA C6, µm,.6 mm Pump: Summit P 68A DGP-6 Mobile Phases: A. :8:.7 acetonitrile:water:tfa B. :66:. acetonitrile:water:tfa Flow:. ml/min Temperature: C Gradient Times: %A %B Injection: Summit ASI, µl Detector: Summit UVDU diode array, UV at 7, 8 and 7 nm. Pseudo-ephedrine µg/ml. Fexofenadine *. Artifacts 9 APPLICATINS - -

34 Environmental Applications Environmental Brodifacoum in Rodent Bait Column: Acclaim C8, µm,.6 mm Pump: Summit P8 HPG/ Mobile Phase: (A) Acetonitrile (B).% conc. phosphoric acid in water m Isocratic A:B 8.:7. Flow:. ml/min Temperature: C Injection: Summit ASI-, µl Detector: Summit UVD U diode array, 6 nm, and spectra nm Sample Prep.: - Mutually saturate ethyl acetate and water for use in the next step - Shake.g of finely ground bait with. ml ethyl acetate,. ml water and. ml acetic acid - Centrifuge to separate the emulsion - Evaporate. ml of ethyl acetate fraction to dryness - Reconstitute in.6 ml acetonitrile - Filter. Brodifacoum isomer. Brodifacoum isomer. Separation of Phthalate Esters in EPA Method 66 Using Acclaim PA 6 Column: Acclaim PA, µm Dimensions:.6 mm Mobile Phase: (A) D.I. H (B) MeH Gradient: Time (min) % B Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: UV, nm ( µg/ml each). Dimethyl phthalate. Diethyl phthalate. Bis(-ethylhexyl) phthalate. Di-n-butyl phthalate. Di-n-octyl phthalate 6. Butyl benzyl phthalate 7 LC Separation of Nitrosamines in EPA 87 Using Acclaim PA Column: Acclaim PA, µm Dimensions:.6 mm Mobile Phase: (A) D.I. H (B) Acetonitrile Gradient: Time (min) % B 9 9 Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: UV, nm ( µg/ml). N-Nitroso-dimethylamine. N-Nitroso-morpholine. N-Nitroso-methylethylamine. N-Nitroso-pyrrolidine. N-Nitroso-diethylamine 6. N-Nitroso-piperidine 7. N-Nitroso-dipropylamine 8. N-Nitroso-dibutylamine 9. N-Nitroso-diphenylamine Note: This gradient will also resolve benzidines. 9a. Phenol Standards by EPA Method 6 on Acclaim PA Note: The lower ph provided by methanesulfonic acid (compared to acetic acid in the standard method) gives optimum separation for the Acclaim PA column. Column: Acclaim PA, µm Dimensions:.6 mm Mobile Phase: (A). mm MSA in H (B) Methanol Gradient: Time (min) % B Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: UV, 8 nm ( µg/ml each). Phenol. -Chlorophenol. -Nitrophenol. -Nitrophenol.,-Dimethylphenol 6. -Chloro--methylphenol 7.,-Dichlorophenol 8.,-Dinitrophenol 9. -Methyl-,6-dichlorophenol.,,6-Trichlorophenol. Pentachlorophenol 6 APPLICATINS LC/MS Detection of Herbicides on Acclaim C8 Column: Acclaim C8, µm Dimensions:. mm. Mobile Phase: 7/% methanol/.% formic acid Flow Rate:. ml/min Detection: AQA MS, +ESI, V, C, SIM as indicated µa m/z mass injected. Monuron 99 ng. Atrazine 6.. Propazine. Diuron.. Linuron Benzidines by EPA Method 6 on Acclaim C8 with Electrochemical Detection Column: Acclaim C8, µm Dimensions:. mm Mobile Phase: / acetonitrile/ mm NaAc, ph.7 Flow Rate:.7 ml/min Inj. Volume: µl Detection: Glassy carbon electrode, 8 mv ( µg/ml) Peaks. Benzidine..'-Dimethylbenzidine..'-Dichlorobenzidine Note: Scaling down the column size and flow rate produces a superproportional increase in sensitivity in the electrochemical detector. - -

35 Environmental Applications Improved Separation of Carbonyl-DNPH Derivatives by CARB Method Using Coupled Acclaim PA and Acclaim C8 Columns Columns: Acclaim PA, µm,.6 mm, coupled with Acclaim C8, µm,.6 mm Mobile Phase: (A) H (B) CH CN Gradient: Time (min) % B. 9 9 Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: UV, 6 nm (. µg/ml each) DNPH derivatives of. Formaldehyde. Acetaldehyde. Acrolein. Acetone. Propionaldehyde 6. Crotonaldehyde 7. Methacrolein 8. Butyraldehyde 9. Methylethylketone. Benzaldehyde. Valeraldehyde. m-tolualdehyde. Hexaldehyde Background Information: LC/MS of insecticides, ng each on column. verlay of simultaneous single-ion chromatograms. Column: Acclaim C8, µm,. mm Mobile Phase: (A) MeH (B) mm NH HC, ph. Gradient: Time %A %B Flow Inj. Volume: µl Sample: ppm stock diluted in Eluent B, Counts Carbamate Insecticides in EPA. on Acclaim C8 with MS Detection Detection: MSQ MS +ESI MS Settings: Probe C Needle. kv Cone voltage set per ion m/z. Aldicarb sulfoxide 7. Aldicarb sulfone. xamyl 7. Methomyl 6. -Hydroxy carbofuran 6. Aldicarb 8 7. Unknown 8 8. Propoxur Carbofuran. Carbaryl. Unknown 69. Methiocarb Notes: () No conventional C8 column can produce equivalent resolution. () CARB = California Air Resource Board m Polynuclear Aromatic Hydrocarbon Standards by EPA Method 8 on Acclaim PA Column: Acclaim PA, µm Dimensions:.6 mm Mobile phase: (A) CH CN (B) D.I. H Gradient: Hold A/B (/) for min, A/B (/) to A/B (8/) in min, Hold A/B (8/) for min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: UV, nm. Naphthalene. Acenaphthylene. Acenaphthene. Fluorene. Phenanthrene 6. Anthracene 7. Fluoranthene 8. Pyrene 9. Benzo[a] anthracene. Chysene. Benzo[b] fluoranthene. Benzo[k] fluoranthene. Benzo[a] pyrene. Dibenz[a,h] anthracene. Benzo[g,h,i] perylene 6. Indeno[,,-cd] pyrene 9 6 m Thirteen Carbonyl-DNPH Derivatives by CARB Method on Acclaim C8 Column: Acclaim C8, µm Dimensions:. mm Mobile Phase: % acetonitrile at 7 min, gradient to % acetonitrile at min Flow Rate:. ml/min Inj. Volume: µl Detection: UV, 6 nm Peaks (DNPH and carbonyl derivatives). µg/ml:.,-dnph. Formaldehyde-DNPH. Acetaldehyde-DNPH. Acrolein-DNPH. Acetone-DNPH 6. Propionaldehyde-DNPH 7. Crotonaldehyde-DNPH 8. Methacrolein-DNPH 9. -Butanone-DNPH. Butyraldehyde-DNPH. Benzaldehyde-DNPH. Valeraldehyde-DNPH. m-tolualdehyde-dnph. Hexaldehyde-DNPH Note: CARB = California Air Resource Board APPLICATINS - -

36 Environmental Applications. Column: Acclaim PA, µm 9 Dimensions:.6 mm Mobile Phase: (A) D. I. H (B) CH CN Carbamate Insecticide Standards in EPA. on Acclaim PA Gradient: Time (min) % B 7 Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: UV, nm ( µg/ml each). Aldicarb sulfoxide. Aldicarb sulfone. xamyl. Methomyl. -Hydroxycarbofuran 6. Aldicarb 7. Propoxur 8. Carbofuran 9. Carbaryl. Methiocarb,, Counts Sensitive Detection of Triazine Herbicides on Acclaim C8 with MS Detection Background Information: Seven herbicides at ng on column. Seven simultaneous single-ion chromatograms optimized for positive identification. Column: Acclaim C8, µm Dimensions:. mm Mobile Phase: % Acetonitrile 6% Ammonium acetate, ph Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: MSQ MS ESI: Positive Probe: C Needle:. kv Cone Voltage: Set per ion Peaks m/z. Simazine. Atrazine 6. Prometon 6. Ametryn 8. Propazine 6. Prometryn 7. Terbutryn LC Separation of Benzidines in EPA 87 Using Acclaim PA Column: Acclaim PA, µm Dimensions:.6 mm Mobile Phase: (A) D.I. H (B) Acetonitrile Gradient: Time (min) % B 9 9 Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: UV, nm ( µg/ml). Benzidine Note: This gradient will also resolve N-nitrosamines..'-Dimethylbenzidine..'-Dichlorobenzidine 9b. Four Antibiotics in Animal Feed on Acclaim C8 6 8 Column: Acclaim C8, µm,.6 mm Pump: GP microbore Mobile Phase A: mm formic acid Mobile Phase B: Acetontirile Gradient Times: %A: 8 %B: Flow:. ml/min Temperature: C Injection: µl Detector: UV, 8 nm ( µg/ml each). Sulfamerazine. xytetracycline. Chloramphenicol. Sulfadimethoxine 66 APPLICATINS EPA Method 8 Using Acclaim Explosives E Column Column: Acclaim Explosives E, mm,.6 mm Mobile Phase: /7 v/v MeH/H Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: UV, nm. HMX. RDX.,,-Trinitrobenzene.,-Dinitrobenzene. Nitrobenzene 6. Tetryl 7.,,6-Trinitrotoluene 8. -Amino-,6-Dinitrotoluene 9. -Amino-,6-Dinitrotoluene.,6-Dinitrotoluene.,-Dinitrotoluene. -Nitrotoluene. -Nitrotoluene. -Nitrotoluene 78 EPA Method 8 Using Acclaim Explosives E Column Column: Acclaim Explosives E, mm,.6 mm Mobile Phase: / v/v MeH/H Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: UV, nm (EPA 8 mix, ppm each). HMX. RDX.,,-Trinitrobenzene.,-Dinitrobenzene. Nitrobenzene 6.,,6-Trinitrotoluene 7. Tetryl 8.,6-Dinitrotoluene 9.,-Dinitrotoluene. -Nitrotoluene. -Nitrotoluene. -Nitrotoluene. -Amino-,6-Dinitrotoluene. -Amino-,6-Dinitrotoluene

37 Food and Beverage Applications Food and Beverage m Carotenoids in Serum on Acclaim PA Spectrum library match Column: Acclaim PA C6, µm,.6 mm Pump: Summit P8 HPG/ Mobile Phase: (A) Water (B) :: methanol:acetonitrile:isopropanol Gradient: Time %A %B 9 9 Flow:. ml/min Temperature: C Injection: Summit ASI-, µl Detector: Summit UVD U diode array, nm and spectra 9 nm Sample: Extract of bovine serum, fortified with lycopene equivalent to ng/ml. Unidentified carotenoid. trans-lycopene. trans-β-carotene. cis-β-carotene Pigments in Turmeric on Acclaim PA 9 Column: Acclaim PA, µm,.6 mm Pump: Summit P68 DGP Mobile Phase: (A) mm phosphoric acid m (B) Methanol Isocratic: 78% B Flow:. ml/min Temperature: Summit TCC-, C Injection: Summit ASI-, µl Detector: Summit UVDU diode array, 8 nm, and spectra 9 nm. Curcumin. Demethoxycurcumin Sample Preparation:. Bis-demethoxycurcumin Extraction solvent: 7% ethanol, 9.% water,.% phosphoric acid Extract mg of ground turmeric with ml of solvent in a capped vial at 6 C for min Centrifuge Dilute in 7% methanol Filter 69 m Fat-Soluble Vitamin Standards on Acclaim C Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: 9/ v/v acetonitrile/methanol Flow Rate:. ml/min Temp.: C Inj. Volume: µl Detection: UV, 8 nm. All-trans retinol (Vitamin A). mg/ml. δ-tocopherol (impurity) -. Ergocalciferol (Vitamin D ).. Cholecalciferol (Vitamin D ).6. β-tocopherol (impurity) - 6. α-tocopherol (Vitamin E) m Amino Acids in Vitamin Premix on Acclaim A Column: Acclaim A, µm Dimensions: mm Mobile Phase: mm Na S adjusted to ph. with methanesulfonic acid Flow Rate:.6 ml/min Temperature: C Inj. Volume: µl Detection: UV, nm. Lysine. Glutamine. Valine. Ascorbic acid. Isoleucine 6. Leucine 7. Unknown 8. Unknown.7 rganic Acids in range Juice on Acclaim A Column: Acclaim A, µm Dimensions: mm Mobile Phase:. M Na S, ph.68 (adjusted with methanesulfonic acid) Temperature: C Flow Rate:.6 ml/min Inj. Volume: µl Detection: UV, nm Sample Prep.: nguard II P, diluted with D.I. H. Malic acid. Ascorbic acid (Vitamin C). Citric acid. Fumaric acid. rganic Acids in White Wine on Acclaim A Column: Acclaim A, µm Dimensions: mm Mobile Phase:. M Na S, ph.68 (adjusted with methanesulfonic acid) Temperature: C Flow Rate:.6 ml/min Inj. Volume: µl Detection: UV, nm Sample prep: nguard II P. Tartaric acid. Malic acid. Lactic acid. Citric acid. Succinic acid APPLICATINS - 7 -

38 Food and Beverage Applications m Assay for Water-Soluble Vitamins in Vitamin Tablets Using Acclaim PA nm 6 nm 6 nm 8 nm 7 ne-a-day is a registered trademark of Bayer Corp. Column: Acclaim PA, µm Dimensions:.6 mm Mobile Phase: Phosphate ph., gradient % CH CN Detection: UVD at listed wavelengths Sample: ne-a-day brand "Active Formula" Sample Prep.:. Grind a mg tablet. Extract mg with ml of ph. buffer. Sonicate min. Filter. Inject µl. Thiamine. Ascorbic acid. Pyridoxine. Niacinamide. Pantothenic acid 6. Folic acid 7. Vitamin B 8. Riboflavin 9 Nitrofuran Antibiotic Residues in Animal Feed on Acclaim C8 m 6 8 Reference: R.J. McCracken, D.G. Kennedy; J. Chromatogr. A, 997, 77, 9. mg/kg mg/kg Control Column: Acclaim C8, µm,.6 mm Pump: Summit P68A DGP-6 Mobile Phase: Acetonitrile: mm ammonium acetate ph., :8 v/v; isocratic Flow:. ml/min Temperature: C Injection: Summit ASI, µl Detector: Summit UVDU diode array, 6 nm, and spectra nm Sample Preparation:.. g guinea pig feed in a ml centrifuge tube. Add 9 ml water and let stand min.. Add ml methanol:acetonitrile : and extract for min.. Pass through cleanup cartridge containing.7g of neutral alumina; discard first.7 ml, retain next. ml. Nitrofurazone. Nitrofurantoin. Furazolidone. Furaltadone 8,, Counts Sensitive Detection of Triazine Herbicides on Acclaim C8 with MS Detection Background Information: Seven herbicides at ng on column. Seven simultaneous single-ion chromatograms optimized for positive identification. Column: Acclaim C8, µm Dimensions:. mm Mobile Phase: % Acetonitrile 6% Ammonium acetate, ph Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: MSQ MS ESI: Positive Probe: C Needle:. kv Cone Voltage: Set per ion Peaks m/z. Simazine. Atrazine 6. Prometon 6. Ametryn 8. Propazine 6. Prometryn 7. Terbutryn m Fat-Soluble Vitamins and Carotenoids in a Vitamin Tablet 7 nm nm 8 nm Sample Preparation: Finely grind a weighed tablet. Weigh an aliquot of about / tablet ( mg) into a screw-cap vial. Partition between. ml water and. ml ethyl acetate with vigorous shaking. Warm to 6 C for min. Shake vigorously again for. min. Centrifuge to separate phases. 6 7 Column: Acclaim PA C6, µm,.6 mm Pump: Summit P68A DGP-6 Mobile Phase: (A) Water (B) :: methanol:acetonitrile: isopropanol Gradient: Times %A %B 9 9 Flow:. ml/min Temperature: C Injection: Summit ASI-, µl Detector: Summit UVD U diode array, 7, and 8 nm, and spectra 9 nm. trans-retinol acetate. trans-lutein. α-tocopherol acetate. trans-lycopene. cis-lycopene 6. trans-β-carotene 7. cis-β-carotene 786 APPLICATINS - 8 -

39 Food and Beverage Applications. Parabens on Acclaim PA 6 8 Column: Acclaim PA, µm Dimensions:.6 mm Mobile Phase: CH CN/H v/v 6/ Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: UV, nm (. mg/ml each). Methylparaben. Ethylparaben. Propylparaben. Butylparaben 6 m *btained from Am. Soc. Brewing Chemists Hop Extract on Acclaim C8 Column: Acclaim C8, µm,.6 mm Pump: Summit P68A DGP-6 Mobile Phase: Methanol, water, phosphoric acid; 8:7:. (v/v/v) Flow:. ml/min Temperature: TCC- thermostat, C Injection: ASI autosampler, µl Detector: UVD-U, UV nm; spectra nm Standard*: ICE- certified hop extract, 986 µg/ml in acidified methanol. Cohumulone µg/ml. Humulone + adhumulone. Colupulone 7. Lupulone + adlupulone Water-Soluble Vitamins Using Acclaim PA Determination of Quinine in Tonic Water Using Acclaim PA Column: Acclaim PA, µm Dimensions:.6 mm Mobile Phase: (A) CH CN (B) mm phosphate buffer, ph. Gradient: Hold B for min B to A/B (/6) in 7 min then to A/B (7/) in min Hold A/B (7/) for min Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: UV, nm. Thiamine. µg/ml. Ascorbic acid 6. Pyridoxine.. Niacinamide. Pantothenic acid 6. Vitamin B. 7. Folic acid 8. Riboflavin 9. Biotin A. Tonic Water B. Quinine standard, µg/ml Reference: U.S. Pharmacopeia, nd Ed., 99, 9. Column: Acclaim PA C6, µm,.6 mm Pump: GP microbore Mobile Phase: (A) 896 ml water, ml acetonitrile,. ml methanesulfonic acid,.8 ml acetic acid,. ml diethylamine (B) 7: v:v acetonitrile:water Gradient: Time...6. %A %B Flow:. ml/min Temperature: C Injection: µl Detector: AD, UV nm Sample Prep.: Filter. Cinchonidine. Quinine. Dihydroquinine. Benzoic acid 8 Determination of Energy Drink Ingredients with Acclaim PA 6 m nm 6 nm 6 nm 8 nm Sample Prep: Column: Acclaim PA, µm,.6 mm Pump: Summit P68 DGP6 Mobile Phases: (A) Acetonitrile (B) mm phosphate buffer, ph.8 (C) mm phosphate buffer, ph. Gradient: Times.. 6. %A %B %C 6 6 Flow:. ml/min Temperature: C Injection: ASI- autosampler, µl Detector: UVD U, UV, 6, 6, 8 nm; spectra nm Filter. Ascorbic acid. Pyridoxine. Niacinamide. Pantothenic acid. Caffeine 6. Riboflavin 7. Sorbic acid 8. Benzoic acid 78 m B A 6 8 Sample Preparation: Finely grind dried peppers Take an aliquot of about mg Partition between. ml water and 6. ml ethyl acetate with vigorous shaking for min Centrifuge to separate phases Evaporate. ml of ethyl acetate phase and reconstitute in. ml ethanol Sudan Dyes in Paprika Column: Acclaim C8, µm,.6 mm Pump: Summit P68 DGP Mobile Phase: (A) Water (B) : tetrahydrofuran:methanol Gradient: Times %A 8 %B 8 9 Flow:. ml/min Temperature: C Injection: Summit ASI-, µl Detector: Summit UVDU diode array, nm, and spectra 9 nm Samples: (A) Control paprika without artificial coloring (B) Paprika spiked with approx. mg/kg of each dye. Sudan I. Sudan II. Sudan III. Sudan IV. il Red ( peaks) (Natural pigments 8- min.) 787 APPLICATINS - 9 -

40 Food and Beverage Applications Determination of Soft Drink Ingredients with Acclaim A Tetracyclines in Pork on Acclaim PA 97 m A B C D Note: For alternate calibration mixes, use ascorbic acid instead of erythorbic acid, or use saccharin instead of aspartame. Minute-Maid and Diet Coke are registered trademarks of Coca-Cola. Mountain Dew is a registered trademark of Pepsico. Column: Acclaim A, µm,. mm Pump: Summit P8 HPG/ Mobile Phase: (A). g/l Na S +. ml/l CH S H in water (B) Methanol Gradient: Time %A %B Flow:.8 ml/min Temperature: C Injection: ASI- autosampler, µl Detector: UVD U, UV nm; optional, 6 nm Samples: (A) Minute-Maid range Soda (B) Diet Coke (C) Mountain Dew (D) Standards Sample Prep.: Filter and dilute with mobile phase A. Erythorbic acid µg/ml. Citric acid. Acesulfame-K. Aspartylphenylalanine. Caffeine 6. Aspartame 7. Sorbate potassium 8. Benzoic acid 8 m A B C 6 7 Sample Prep.: -. g pork muscle tissue - Grind with mortar and pestle in sequence with. g of citric acid. ml of.6 M HN. ml of methanol. ml of water - Centrifuge Reference: Farrington et. al., Food Additives and Contaminants, 99, Vol. 8, No., -6. Column: Acclaim PA C8, µm,.6 mm Pump: Summit P8 HPG/ Mobile Phase: (A) mm H P,. mm Na P 7 (B) Acetonitrile Gradient: Time.. 7. %A 8 8 %B 8 8 Flow:. ml/min Temperature: C Injection: ASI- autosampler, 6 µl Detector: UVD U, UV at 6 nm Samples: (A) Spiked pork muscle,. µg/g each (B) Control pork (C) Tetracycline standards,. µg/ml in water. xytetracycline. Tetracycline. Chlortetracycline. Doxycycline m Fat-Soluble Vitamins and Carotenoids in Dry Pet Food nm 7 nm Sample Preparation: Extraction by AAC fficial Method g of finely ground feed Add ml of hexane-acetone-ethanol-toluene, :7:6:7, v/v/v/v Add ml of % KH in methanol Saponify at 6 C for min Cool for hr Add ml hexane Dilute to ml with % sodium sulfate solution Shake vigorously For HPLC, evaporate. ml of hexane phase and reconstitute in. ml of acetone Column: Acclaim PA C6, µm,.6 mm Pump: Summit P68A DGP-6 Mobile Phase: (A) Water (B) :: methanol:acetonitrile: isopropanol Gradient Times: %A %B 9 9 Flow:. ml/min Temperature: C Injection: Summit ASI-, µl Detector: Summit UVD U diode array, 7, and 8 nm, and spectra 9 nm. trans-retinol. trans-lutein. trans-β-carotene 66 Glutamate in Beer by Automated Precolumn PA Derivatization y =.97x +.8 R = Added Glutam ate (µg/ml) m Glutamate Column: Acclaim PA, C6 µm,.6 mm Pump: Summit P68A Mobile Phases: (A) Methanol (B) mm phosphate, ph 7. Gradient Times: %A: 7 7 %B: Flow:. ml/min Temperature: TCC thermostat at C Injection: ASI- autosampler; µl Detector: UVD U; UV at nm and spectra nm Samples: (A) Beer (B) Beer + µg/ml glutamic acid Derivatizing Conditions: Program: (Reagent A) mg/ml o-phthalaldehyde in.m borate buffer, ph. (Reagent B) 6 mg/ml (CH ) NCH CH SH HCl in water 7 µl reagent A + 7 µl reagent B + µl sample, mixed in round-bottom B glass vial Reaction Time: min at room temperature A 6 Fat-Soluble Vitamin Standards Using Acclaim PA Four Aflatoxin Standards on Acclaim C8 APPLICATINS m 8 6 Column: Acclaim PA, µm Dimensions:. mm Mobile Phase: 97/ acetonitrile/nh HC, ph. Flow Rate. ml/min Temperature: C Inj. Volume: µl Detection: UV, 8 nm. Vitamin A acetate. Vitamin D. Vitamin D. Vitamin E acetate. Vitamin E 6. Vitamin K 9 m Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: Isocratic,././ methanol/acetonitrile/water Flow Rate:. ml/min Temperature: C Inj. Volume: µl Detection: UV, 6 nm. G 7. ng. G. B 7.. B - -

41 Food and Beverage Applications Sulfonamide Antibiotic Residues in Milk on Acclaim C8 Antioxidants in Edible ils by AAC 98. on Acclaim C8 C m B * A Column: Acclaim C8, µm,.6 mm Pump: Summit P8 HPG/ Mobile Phase: (A) mm acetic acid adjusted to ph. with NaH (B) Methanol Gradient Times:. %A %B Flow:. ml/min Temperature: C Injection: ASI- autosampler, µl Detector: UVD U, UV at 6 nm with spectral confirmation Sample Prep.: AAC 99.; refer to procedure for important notes Samples: (A) Spiked extract of skim milk,. µg/g each (B) Milk extract (C) Sulfonamide standards,. µg/ml in water. Sulfadiazine (SDZ). Sulfathiazole (STZ). Sulfapyridine (SPD). Sulfamerazine (SMR). Sulfamethazine (SMZ) 6. Sulfachloropyridazine (SCP) 7. Sulfadimethoxine (SDM) 8. Sulfaquinoxzline (SQX) *. Theobromine Note: Most interferences are organic acids that may be moved out of the way by small adjustements to the ph of the mobile phase. Background Information: Nine antioxidants for AAC 98. spiked into wheat germ oil. Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: (A).7% HAc in H (B) //. MeH/acetonitrile/HAc Inj. Volume: µl Detection: UV, 8 nm; diode array confirmation Gradient: Time % A % B Flow m Propyl gallate. THBP. TBHQ. NDGA. BHA 6. Ionox- 7. ctyl gallate 8. BHT 9. Dodecyl gallate 9 8 Note: Ethoxyquin (not shown) has been resolved under the same conditions. 8 9 Extract of Spiked Wheat Germ il Extract of Wheat Germ il Standards, ng each Bitter Principles in Beer on Acclaim C8 Standards m ICS-H ICS-R ICS-T ICS-I 6. Märzen-style beer m Column: Acclaim C8 µm,.6 mm Pump: Summit P68A DGP-6 Mobile Phase: 7% methanol, % water, % phosphoric acid (v/v/v) Flow:. ml/min Temperature: TCC- thermostat, C Injection: ASI autosampler, µl Detector: UVD-U, UV 7 nm; spectra nm Standards*: ICS-I: isohumulones µg/ml ICS-T: tetrahydroisohumulones ICS-R: rho-isohumulones ICS-H: hexahydroisohumulones. Isocohumulone. Mixed isohumulone congeners. Isoadhumulone Sample Preparation: Extraction per ASBC - ml beer + ml N HCl + µl -octanol + ml iso-octane - Shake vigorously - Centrifuge to separate phases - Bitterness Units = A 7 For HPLC - Evaporate dry and reconstitute in mobile phase *btained from Am. Soc. Brewing Chemists 7 APPLICATINS - -

42 Chemical Applications Surfactants mv Separation of Cationic Surfactants Using ELSD Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) Acetonitrile (B). M NH Ac, ph. Gradient: 8% A in min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELSD. Lauryl pyridinium chloride. Lauryldimethylbenzyl ammonium chloride. ctylphenoxyethoxyethyl dimethylbenzyl ammonium chloride. Cetyltrimethylammonium chloride. Cetylpyridinium chloride 8 mv UV, nm ELSD Analysis of a Shampoo Laureth Sulfate Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) Acetonitrile (B). M NH Ac, ph. Gradient: 8% A in min, then hold at 8% A for min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELSD, UV 8 Analysis of a Mouthwash Analysis of a Fabric Softener mv Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) Acetonitrile (B). M NH Ac, ph. Gradient: 8% A in min Temperature: C Flow Rate: ml/min Inj. Volume: µl (direct injection) Detection: ELSD. Sodium sacchrin and benzoate. Domiphen bromide. Cetylpyridinium chloride mv Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) Acetonitrile (B) THF (C). M NH Ac, ph. Gradient: % A/% B/% C to 7% A/ % B/7% C in min, then hold for min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELSD Methyl bis[ethyl(tallowate)]-- hydroxyethyl ammonium methyl sulfate ( ) Analysis of ZNYL FS Fluorosurfactant Separation of Ethoxylated Quats mv Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) Acetonitrile, (B) H Gradient: 7% A in min, then hold at 7% A for min Flow Rate: ml/min Inj. Volume: µl Detection: ELSD R f CH CH (CH CH ) x H mv Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: Acetonitrile/. M NH Ac, ph. v/v /6 Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELSD Sample: Ethoquad 8/ ( ) R f = F(CF CF ) y x = to about y = to about 7 Zonyl FS fluorosurfactant APPLICATINS

43 Chemical Applications Surfactants Analysis of Sodium Lauryl Sulfate Using LC-ESI-MS Analysis of PEG Monoethyl Ether (MW-) Abundance Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: Acetonitrile/. M NH Ac, ph. v/v 7/ Temperature: C Flow Rate: ml/min (. ml/min to mass spectrometer) Inj. Volume: µl Detection: MS (ESI negative, probe C, cone V needle. KV, SIM 6., span. m/z) Peak: Sodium lauryl sulfate ( ppm) mv Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) Acetonitrile (B) H Gradient: % A in min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELSD Abundance Analysis of Triton X- Using LC-ESI-MS 8 6 Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: Acetonitrile/ mm NH Ac, ph. v/v / Temperature: C Flow Rate: ml/min (. ml/min to mass spectrometer) Inj. Volume: µl Detection: MS (ESI positive, probe C, cone V, needle. kv, SIM., 6.,.,., 88.,., 76., 6.6, 66.6, 78.6, 7., 796.6, 8.7, 88.7, 98.7, 97.7, 6.8, 6.7,.7, span. m/z) Peak: C 8 H 7 Triton X- ( ppm) (C HCH ) n H mv Separation of Hydrotrope, Anionic, Cationic, and Amphoteric Surfactants Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) CH CN, (B).6 M NH Ac, ph. Gradient: % to 8% A in min, then hold 8% A for min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELS detector Xylene sulfonate. Laurylpyridinium chloride. Lauryldimethylbenzyl ammonium chloride. ctylphenoxyethoxyethyl dimethylbenzyl ammonium chloride. Myristamidopropyl betaine 6. Cetyltrimethyl ammonium chloride 7. Cetylpyridinium chloride 8. Cetyl betaine 9. Decyl sulfate. Dodecyl sulfate. C -LAS. C -LAS. C -LAS. C -LAS n ~ Triton X UV, nm Dodecylbenzene sulfonate ELSD Analysis of a Laundry Washing Detergent Dodecylbenzene sulfonate and Laureth sulfate Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) Acetonitrile (B). M NH Ac, ph. Gradient: 8% A in min, then hold at 8% A for min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELSD, UV Sample Prep: Dilute with 7% acetonitrile, then filtered through.-µm membrane UV, nm Dodecylbenzene sulfonate ELSD Dodecylbenzene sulfonate and laureth sulfate Analysis of a Dishwashing Liquid Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) Acetonitrile (B). M NH Ac, ph. Gradient: 8% A in min, then hold at 8% A for min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELSD, UV Sample Prep: Dilute with 7% acetonitrile, then filtered through.-µm membrane APPLICATINS

44 Chemical Applications Surfactants Analysis of a Strongly Hydrophilic Hydrotrope Acclaim Surfactant Conventional C8 Dimensions:.6 mm Mobile Phase: Acetonitrile/. M NH Ac, ph. v/v /7 Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: UV, nm Sample: Sodium xylene sulfonate ( ppm) 7 µs Analysis of Ammonium Laureth Sulfate Using Conductivity Detection Column: Acclaim Surfactant, µm Dimension:.6 mm Mobile Phase: Acetonitrile/. M H B and mm NaH v/v / Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: Suppressed CD (AMMS III) Sample: Ammonium laureth sulfate 76 Analysis of NEDL -7 Analysis of Quats Using LC-ESI-MS mv R (CH CH ) n H R = C to C n ~ 7 NEDL -7 Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) Acetonitrile (B) H Gradient: 6% A in min, then hold at 6% A for min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELSD Abundance C C6 C7 C8 Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: Acetonitrile/ mm NH Ac, ph. v/v 6/ Temperature: C Flow Rate: ml/min (. ml/min to mass spectrometer) Injection Vol.: µl Detection: MS (ESI positive, probe C, cone V, needle. kv, SIM., 6., 7., 88., span. m/z) Peak: Benzyl dimethyl hydrogenated tallow ammonium chloride ( ppm) 79 7 APPLICATINS mv Analysis of Toilet Bowl Cleaner. M NH Ac M NH Ac Separation improved by changing mobile phase ionic strength Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) CH CN (B) NH Ac, ph. Gradient: % to 8% A in min, hold for min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELS detector Sample: Toilet bowl cleaner ( dilution). Chloride. PEG 7. Cationic surfactants 8. Nonionic surfactant 797 mv Inorganic Anions, Hydrotropes, Cationic, Nonionic, Amphoteric, and Anionic Surfactants Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) CH CN, (B). M NH Ac, ph. Gradient: % to 8% A in min, then hold 8% A for min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELS detector Chloride. Bromide. Nitrate. Xylene sulfonate. Laurylpyridinium chloride 6. Lauryldimethylbenzylammonium chloride 7. Triton X- 8. Cetyl betaine 9. Decyl sulfate. Dodecyl sulfate. C -LAS. C -LAS. C -LAS. C -LAS

45 Chemical Applications Surfactants Separation of Different Polyethylene Glycols Separation of Quaternary Imidazolinium Compounds mv PEG- PEG- Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) Acetonitrile (B) H Gradient: % A in min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELSD mv -Hydroxyethylbenzyl coco imidazolinium chloride Diethyl heptadecyl imidazolinium ethylsulfate Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) Acetonitrile (B). M NH Ac, ph. Gradient: 8% A in min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: ELSD PEG µs Separation of Cationic Surfactants with Suppressed Conductivity Detection Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) Acetonitrile (B) mm acetic acid Gradient: % A in min Temperature: C Flow Rate: ml/min Inj. Volume: µl Detection: Suppressed CD (AMMS III). Lauryl pyridinium chloride. Lauryldimethylbenzyl ammonium chloride. ctylphenoxyethoxyethyl dimethylbenzyl ammonium chloride. Cetyltrimethylammonium chloride. Cetylpyridinium chloride 86 mv Analysis of PEG- Distearate Column: Acclaim Surfactant, µm Dimensions:.6 mm Mobile Phase: (A) CH CN, (B). M NH Ac, ph. Gradient: 8% A in min, then hold 8% A for min Temperature: C Flow Rate:. ml/min Inj. Volume: µl Detection: ELS detector Sample: PEG- Distearate 796 PE() Ammonium Lauryl Sulfate Dyes µs Column: Acclaim PA, µm Dimension:.6 mm Mobile phase: Acetonitrile/ mm borate buffer v/v /6 Temperature: C Flow Rate: ml/min Inj. Volume: µl ( ppm) Detection: Suppressed conductivity detection (AMMS III) Sample: PE() Ammonium lauryl sulfate m Sudan Dyes on Acclaim C8 Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: //. MeH/acetonitrile/HAc Flow Rate:. ml/min Inj. Volume: µl Detection: UV, 8 nm. Sudan I. Sudan I, impurity. Sudan II. Sudan III 7 6 APPLICATINS - -

46 Bioscience Applications Proteins and Peptides Tryptic Peptide Map of Cytochrome c on Acclaim C8 Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: (A) 9//. H /acetonitrile/tfa (B) /9/. H /acetonitrile/tfa Flow Rate:. ml/min Gradient: % B to % B in min Inj. Volume: µl Detection: UV, nm m Separation of Protein Standards by Micro LC on Acclaim C8 Column: Acclaim C8, µm Dimensions: -µm i.d. cm Mobile Phase: (A) Water..% TFA (B) 8/ CH CN/ water,. % TFA Gradient: 6% B in min Flow Rate: µl/min Detection: UV, nm. Ribonuclease ng. Insulin. Cytochrome c. Myoglobin 9 Tryptic Peptide Map of Bovine Serum Albumin on Acclaim C8 Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: (A) 9//. H /acetonitrile/tfa (B) /9/. H /acetonitrile/tfa Flow Rate:. ml/min Gradient: % B to % B in min Inj. Volume: µl Detection: UV, nm Capillary LC for Peptide Mapping Using Acclaim C8 Column: Acclaim C8, µm Dimensions: 7 µm i.d. cm Mobile Phase: (A) 9/ (v/v) water/acetonitrile,.% TFA (B) /8 (v/v) water/acetonitrile,.% TFA Gradient: 6% B in 6 min Sample: Tryptic digest of BSA, pmol/µl Detection: UV, nm m APPLICATINS - 6 -

47 Bioscience Applications Proteins and Peptides m Cytochrome c from Three Species on Acclaim C8 6 8 Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: (A) 9//. v/v water/acetonitrile/ TFA (B) 9//. v/v acetonitrile/water/ TFA Gradient: Time (min) % A % B 8 8 Flow Rate: Inj. Volume: Detection:. ml/min µl ( µg/µl) UV, nm. Yeast cytochrome c. Equine cytochrome c. Bovine cytochrome c 9 Protein Mixture by Reversed-Phase HPLC on Acclaim C8, m Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: (A) mm MSA in 9/ H /acetonitrile (B) mm MSA in /9 H /acetonitrile Inj. Volume: µl Detection: UV, nm Gradient: Time % A % B Flow. 8. RNAse A. Impurity in carbonic anyhdrase. Impurity in lysozyme. Lysozyme. Impurity in carbonic anhydrase 6. Myoglobin 7. Carbonic anhydrase 7 Comparison of Acid Additives for Peptide Analysis on Acclaim A B C 6 8 Column: Acclaim C8, µm,.6 mm Pump: Summit P68A Mobile Phases: A: 7% acetonitrile v/v B: Water C: Acid stock solution Gradient Times: 8 6 %A %B %C Flow:. ml/min Temperature: TCC thermostat at C Injection: ASI- autosampler; µl Detector: UVD U; UV at nm Acid Additive: A. Phosphoric Acid,.% B. Trifluoroacetic Acid,.% C. Formic Acid,.% Sample: Sigma H6 Peptide Mix. Gly-Tyr. Val-Tyr-Val. Met-Enkephalin. Angiotensin-II. Leu-Enkephalin 6. Peptide Test Mix on Acclaim and PA with Formic Acid A B 6 8 Columns: (A) Acclaim C8, µm,.6 mm (B) Acclaim PA, µm,.6 mm Pump: Summit P68A Mobile Phases: (A) 7% acetonitrile v/v (B) Water (C).% formic acid v/v Gradient Times: 8 6 %A %B %C Flow:. ml/min Temperature: TCC thermostat at C Injection: ASI- autosampler; µl Detector: UVD U; UV at nm Sample: Sigma H6 Peptide Mix. Gly-Tyr. Val-Tyr-Val. Met-Enkephalin. Angiotensin-II. Leu-Enkephalin 66 APPLICATINS - 7 -

48 Natural Products m Alkaloids in Goldenseal Root on Acclaim C8 with ASE Extraction Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: 7/7.% H P in D.I. H / acetonitrile Flow Rate:.8 ml/min Inj. Volume: µl Detection: UV, nm. Hydrastine. Berberine Isoflavones in Red Clover on Acclaim C8 with ASE Extraction 76, m Flavonoids in Green and Black Tea Using Acclaim C8 Green Tea Black Tea 8 6 Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: (A) Acetonitrile (B) mm H P Temperature: C Gradient: Time % A % B Flow Inj. Volume: µl Detection: UV, with PDA spectra. Epigallocatechin. Caffeine. Epicatechin. Epigallocatechin gallate. Unidentified flavonoid 6. Epicatechin gallate m Column: Acclaim C8, µm Dimensions:.6 mm Mobile Phase: (A).% H P in D.I. H (B) Acetonitrile Gradient: % B to 76% B over min Flow rate:. ml/min Inj. Volume: µl Detection: UV, 6 nm. Genistin. nonin. Sissotrin. Formononotin. Biochanin A 769 APPLICATINS - 8 -

49 rganic Acids Separation of Hydroxybenzoic Acids on Acclaim A Resolution of Benzene Polycarboxylic Acids on Acclaim A Column: Acclaim A, µm Dimensions: mm Mobile Phase: /7 v/v CH CN/ mm methanesulfonic acid Temperature: C Flow Rate:.7 ml/min Inj. Volume: µl Detection: UV, nm (8 ppm each).,,-trihydroxybenzoic acid.,-dihydroxybenzoic acid. -Hydroxybenzoic acid.,-dihydroxybenzoic acid. Benzoic acid.7 Column: Acclaim A, µm, Dimensions: mm Mobile Phase:./87. v/v CH CN/ mm methanesulfonic acid Temperature: C Flow Rate:.7 ml/min Inj. Volume: µl Detection: UV, nm ( ppm each).,,-benzenetricarboxylic acid.,-benzenedicarboxylic acid.,,-benzenetricarboxylic acid.,-benzenedicarboxylic acid.,-benzenedicarboxylic acid 8 Acrylic Acid ligomers on Acclaim A Acrylic acid (Aldrich) H Acrylic acid oligomers (Aldrich) H 67 n Column: Acclaim A, µm Dimensions: mm Mobile Phase: (A) CH CN (B). mm methanesulfonic acid Gradient: Hold % A for min, then increase to 9% A in 9 min Temperature: C Flow Rate:.7 ml/min Inj. Volume: µl Detection: UV, nm Sample prep.: Diluted with D.I. H. Monomer. Dimer. Trimer. Tetramer. Pentamer 6. Hexamer 7. Heptamer 9 APPLICATINS - 9 -

50 Lot Qualification Tests Part Four: Appendix Appendix : Lot Qualification Tests for Acclaim-Bonded Silica Column Performance Test, Acclaim C8, C8, PA, PA Polar Selectivity This test protocol is part of the column performance test: column pressure, retention time, efficiency, and asymmetry for phenanthrene are specified for each column. This parameter also is used to monitor batch-to-batch reproducibility as measured by the selectivity of a polar analyte (dimethylphthalate) relative to a nonpolar analyte (phenanthrene). Mobile Phase: 7/ v/v acetonitrile/water Flow Rate:. ml/min Temperature: C Inj. Volume: µl Detection: UV, nm, 6-mm path length. Uracil. mg/ml Metal Activity. Dimethylphthalate.7 µl/ml. Phenanthrene. mg/ml, -Bipyridyl chelates with metals whereas, -bipyridyl does not form chelation complexes. If metals are present, the, isomer elutes with a high asymmetry factor (tailing). The peak asymmetry ratio for the two analytes should be approximately. if no metal contamination is present. Mobile Phase: / v/v methanol/water Flow Rate:. ml/min Temperature: C Inj. Volume: µl Detection: UV, nm., -Bipyridyl ng/µl., -Bipyridyl ng/µl Base Asymmetry Basic compounds exhibit poor peak shape when the silica surface is incompletely covered during the bonding process, or when the silica substrate is inadequately prepared. This test monitors surface coverage with an application-based analysis. Mobile Phase: 8/ v/v methanol/ mm phosphate, ph 6. Flow Rate:. ml/min Temperature: C In. Volume: µl Detection: UV, nm. Uracil. mg/ml. Propranolol.6 mg/ml. Toluene.8 µl/ml. Amitriptyline. mg/ml Hydrophobic Steric Selectivity This parameter is used to monitor batch-to-batch bonding reproducibility as measured by the relative selectivity of two differently shaped aromatic analytes: o-terphenyl and triphenylene. This test is used to characterize all the C8 and polar-embedded phases. Mobile Phase: 9/ v/v methanol/water Flow Rate:. ml/min Temperature: C Inj. Volume: µl Detection: UV, nm. Toluene µl/ml. Phenanthrene. mg/ml. o-terphenyl. mg/ml. Triphenylene. mg/ml APPENDIX Acid Asymmetry This test is used to characterize polar-embedded phases. Acidic compounds exhibit poor peak shape when the bonded phase has basic sites due to degradation or contamination. Mobile Phase: /6 v/v acetonitrile/.% TFA in water Flow Rate:. ml/min Temperature: C Inj. Volume µl Detection: UV, nm. -Hydroxybenzoic acid ng/µl. Benzoic acid 7 ng/µl. Ethyl -hydroxybenzoate ng/µl Some application-specific tests are used for lot validation. Read about these tests in the chapters for Acclaim and Acclaim A. - -

51 Lot Qualification Tests Column Performance Test, Acclaim Surfactant Isocratic Surfactant Selectivity Each column is tested under these conditions to guarantee that the column is well manufactured. Column pressure, efficiency, asymmetry and retention time of propylbenzene are specified for each column. This test also is used to monitor batch-to-batch reproducibility as measured by the relative retention of an anionic surfactant to a neutral marker. Mobile Phase: : (v/v) acetonitrile :.M NH Ac, ph. Flow Rate:. ml/min Temperature: C Inj. Volume: µl Detection: UV, nm. Uracil. mg/ml. -(-octylphenoxy)ethoxyethyldimethylbenzyl ammonium chloride. mg/ml. Sodium p-toluenesulfonate. mg/ml. Propylbenzene. mg/ml Column Performance Test, Acclaim Explosives E and E Each column is tested under these conditions to guarantee that the column is well manufactured. Column pressure, efficiency, asymmetry, and retention time of naphthalene are specified for each column. Mobile Phase: 7/ v/v acetonitrile/water Flow Rate:. ml/min Temperature: C Inj. Volume: µl Detection: UV, nm. Uracil. mg/ml. Naphthalene. mg/ml Gradient Surfactant Resolution An application-based test for the resolution of anionic, nonionic and cationic surfactants. Mobile Phase A: Acetonitrile Mobile Phase B:. M Ammonium acetate, ph. Time %A %B Flow Rate:. ml/min Temperature: C Inj. Volume: µl Detection: Evaporative Light Scattering Analytes:. Laurylpyridinium chloride. mg/ml. Lauryldimethylbenzyl ammonium chloride. mg/ml. Triton X-. mg/ml. Sodium decylsulfate. mg/ml. Sodium dodecylsulfate. mg/ml APPENDIX - -

52 Topical Index Appendix : Topical Index Acclaim C8 -, 7, -,, 9-, 8, C8 -, 7,, -,,, C8 -, 7,, 6-7,, 6-7, PA -, 7, -, 8-, 9-, -, PA -, 7,, -, -, 9, 7, PolarAdvantage, see Acclaim PA and PA A -6, -,, 7,, 9 Surfactant -6, -, -, Explosives -6, 6-7, 6, Acid Aromatic, 9 Asymmetry, 9, Asymmetry test Hydrophilic rganic, 6, -, 7 Alkaline Buffers 9, Amitriptyline, 9, Antibiotics Asymmetry,, 6, 9,,,, 7 Acid, 9,,, Base, 9,, Base Asymmetry, 9, Asymmetry test, Basic Buffers 9, Drugs, 9- Beverages, 7- Bipyridyl, Buffers 9 Capillary,, 7, 9 CARB Method Carbon Load 8 Column Diameter 8 Format 6, 8 Length 8 Corrosion,, 6, 8,, Cytochrome c 6, 6-7 Detection limit 8 De-wetting 8, Efficiency 7,,, 6, 9,,,, 7, ELSD, -, Endcapping, 8,,, 6, 8, Environmental -6 EPA Methods , , 6 Evaporative Light Scattering Detection (ELSD), -, Explosives, 6, 6-7, 6 Flow rate 8 Food and Beverages 7- Guard cartridge,, 7, 9,,,, 7 Hydrolytic stability, 9, 8, - Hydrophilic 8, Hydrophobic Steric selectivity test,, Hydrophobicity 6 Ion Exchange 6 k, Loading Capacity 8 Lot Qualification (see lot validation) Lot Validation,,, 6, 9,,,, 7, - LC-MS, 6, Compatibility,, 7, 9 Metal Activity test,, 6, 9,, Contamination Methanesulfonic acid Mobile Phase 9 Natural products 8 Nitramine 6, 6 Nitroaromatic 6, 6 Normal-phase 8 ctadecyldimethylsilane, 6 ctyldimethylsilane rdering information inside front cover,,, 7, 9,,,, 7 rganic acids -,, 7, 9 PA -, 7, -, 8-, 9-, -, PA -, 7,, -, -, 9, 7, Particle Size, 7 PEEK Peptide, 6, 6-7 ph, 9,,, 6, 8,,,, - Pharmaceutical 9- Phenanthrene Polar Embedded -, 8- Selectivity Test PolarAdvantage, see PA and PA Polarity index, Pore Size, 7 Pressure 7, 8 Protein, 6-7, 6-7 Resolution Reversed-phase 6 Selectivity, 6-7,, 8,,,, 6 Base 7 Ethylbenzene/toluene 7 Peptide 6 Polar Shape 6-7 Steric Tripheylene/o-terphenyl 6-7, Silanol 6, 8,,,, 8 Silica Substrate Suppressed Conductivity Detection, Surface Area, 7,,, 6, 8,,,, 7 Surfactants 6, -, -, Tryptic digest 6, 6 USP Code,,, 6 Validation (see lot Validation) Vitamins 7- Warranty inside front cover Wetting,, 8,, APPENDIX - -

53 Part Number Index Appendix : Part Number Index Part No. Acclaim Type Size Page Part No. Acclaim Type Size Page Part No. Acclaim Type Size Page 9, µm, C8. mm 9, µm, C8. mm 9, µm, C8. mm 9, µm, C8.6 mm 96, µm, C8.6 mm 97, µm, C8.6 mm 98, µm, C8. mm 99, µm, C8. mm 9, µm, C8. mm 9, µm, C8.6 mm 9, µm, C8.6 mm 9, µm, C8.6 mm 9, µm, C8. mm 9, µm, C8. mm 96, µm, C8. mm 97, µm, C8. mm 98, µm, C8.6 mm 99, µm, C8.6 mm 9, µm, C8.6 mm 9, µm, C8.6 mm 9, µm, C8. mm 9, µm, C8. mm 9, µm, C8. mm 9, µm, C8. mm 96, µm, C8.6 mm 97, µm, C8.6 mm 98, µm, C8.6 mm 99, µm, C8.6 mm 96, µm, C8. mm 97, µm, C8 mm 98, µm, C8. mm 99, µm, C8 mm 96 Guard Cartridge holder,, 7,9,,, 7 97 Guard to analyical coupler,, 7,9,,, 7 96 Guard holder & coupler kit,, 7,9,,,, 7 66, µm, C8. mm 7 66, µm, C8. mm 7 66, µm, C8.6 mm 7 666, µm, C8.6 mm 7 69, µm, C8. mm 7 69, µm, C8 mm 7 66 PA, µm, C6. mm 9 67 PA, µm, C6. mm 9 68 PA, µm, C6.6 mm 9 69 PA, µm, C6.6 mm 9 6 PA, µm, C6.6 mm 9 6 PA, µm, C6.6 mm 9 6 PA, µm, C6 mm 9 6 PA, µm, C6. mm 9 69 A, µm. mm 69 A, µm. mm 69 A, µm Guard. mm 687 PA, -µm. mm 689 PA, -µm.6 mm 69 PA, -µm.6 mm 69 PA, µm Guard. mm (pack of ) 69 PA, µm Guard. mm (pack of ) 697 PA, -µm.6 mm 699 PA, -µm.6 mm 6 Surfactant µm.6 mm 6 Surfactant µm.6 mm 6 Surfactant µm Guard. mm (pack of ) 6 Expolsives.6 mm E 7 6 Expolsives µm Guard. mm E (pack of ) 7 67 Expolsives µm Guard. mm E (pack of ) 7 69 Expolsives µm.6 mm E 7 6 Expolsives µm E and E kit 7 6, µm, C8. mm 6, µm, C8. mm 6, µm, C8. mm 6, µm, C8 µm mm 6, µm, C8 µm mm 6, µm, C8 µm mm 66, µm, C8 7 µm mm 67, µm, C8 7 µm mm 68, µm, C8 7 µm mm 6, µm, C8. mm 6, µm, C8. mm 66, µm, C8 µm mm 67, µm, C8 µm mm 68, µm, C8 7 µm mm 69, µm, C8 7 µm mm 6, µm, C8. mm 6, µm, C8. mm 6, µm, C8. mm 6, µm, C8 µm mm 6, µm, C8 µm mm 6, µm, C8 µm mm 66, µm, C8 7 µm mm 67, µm, C8 7 µm mm 68, µm, C8 7 µm mm 69, µm, C8. mm 7 6, µm, C8. mm 7 6, µm, C8 µm mm 7 6, µm, C8 µm mm 7 6, µm, C8 7 µm mm 7 6, µm, C8 7 µm mm 7 6, µm, C8. mm 6, µm, C8. mm 66, µm, C8 µm mm 67, µm, C8 µm mm 68, µm, C8 7 µm mm 69, µm, C8 7 µm mm 6 PA, µm, C6. mm 9 6 PA, µm, C6. mm 9 6 PA, µm, C6 µm mm 9 6 PA, µm, C6 µm mm 9 6 PA, µm, C6 7 µm mm 9 6 PA, µm, C6 7 µm mm 9 66 PA, µm, C6. mm 9 67 PA, µm, C6. mm 9 68 PA, µm, C6. mm 9 69 PA, µm, C6 µm mm 9 6 PA, µm, C6 µm mm 9 6 PA, µm, C6 µm mm 9 6 PA, µm, C6 7 µm mm 9 6 PA, µm, C6 7 µm mm 9 6 PA, µm, C6 7 µm mm 9 66, µm, C8. mm 66, µm, C8. mm 66, µm, C8 8 µm mm 66, µm, C8 µm mm 66, µm, C8 µm mm 666, µm, C8 µm mm 6 PA, µm, C6 µm mm 9 6, µm, C8 µm. mm 66, µm, C8 µm. mm 6 PA, µm, C6 µm. mm 9 6, µm, C8. mm 6, µm, C8. mm 6, µm, C8 8 µm mm 6, µm, C8 µm mm 6, µm, C8 µm mm 66, µm, C8 µm mm 6 PA, µm, C6. mm 9 6 PA, µm, C6. mm 9 6 PA, µm, C6 8 µm mm 9 66 PA, µm, C6 µm mm 9 7 PA, µm, C6 µm mm 9 APPENDIX - -

54 Analyte Index Appendix : Analyte Index Acebutolol..., 9, Benzyl alcohol... 9 Cytochrome-c... 6, 7 Fluoranthene... APPENDIX Acenaphthalene... Acenaphthene... Acesulfame-K... Acetaldehyde-DNPH... Acetaminophen..., Acetic acid... Acetone-DNPH... cis-aconitic acid... trans-aconitic acid... Acrolein-DNPH... Acrylic acid... 9 Acrylic acid dimer... 9 Adhumulone... 9 Adlupulone... 9 Aflatoxin B... Aflatoxin B... Aflatoxin G... Aflatoxin G... Aldicarb..., 6 Aldicarb sulfone..., 6 Aldicarb sulfoxide..., 6 Ametryn Amino-,6-dinitrotoluene...6, 7, 6 -Amino-,6-dinitrotoluene...6, 7, 6 Amitriptyline..., 9,,, Amobarbital... Angiotensin-II... 7 Aniline... 7 Anthracene... Antibiotics...,, 6, 8,, Antidepressants..., 9,,, Antioxidants... Ascorbic acid (Vitamin C)...7, 8, 9 Aspartame... 8, Aspartylphenylalanine... 8, Aspartyl-phenylalanine methyl ester.8, Atenolol..., 9 Atrazine...,, 6 Barbital... Benzaldehyde... Benzaldehyde-DNPH... Benzene,,-tricarboxylic acid... 9 Benzene,-dicarboxylic acid... 9 Benzene,,-tricarboxylic acid... 9 Benzene,-dicarboxylic acid... 9 Benzene,-dicarboxylic acid... 9 Benzidine..., 6 Benzil... Benzo[a]anthracene... Benzo[a]pyrene... Benzo[b]fluoranthene... Benzo[g,h,i]perylene... Benzo[k]fluoranthene... Benzoic acid..., 9,,, Benzoin... Benzophenone... Benzophenone-... -Benzyl-,6-dioxo--piperazine acetic acid... 8 Benzyldimethyl hydrogenated tallow ammonium chloride... Berberine... 8 Biochanin A... 8 Biotin... 8, 9, -Bipyridyl...,, -Bipyridyl..., Bis(-ethylhexyl)phthalate... Bisdemethoxycurcumin... 7 Bovine serum albumin tryptic digest... 6 Brodifacoum... Bromide..., Budesonide... Butabarbital... -Butanone-DNPH... -Butylbenzoic acid..., Butyl paraben... 9 Butylated hydroxyanisole (BHA)... Butylated hydroxytoluene (BHT)... Butylbenzylphthalate... Butyraldehyde-DNPH... Butyric acid... C-LAS...,, C-LAS...,, C-LAS...,, C-LAS...,, Caffeine...9,, 8 Caproic acid... Carbamate insecticides..., 6 Carbaryl..., 6 Carbofuran..., 6 Carbonic anhydrase... 6, 7 beta-carotene...7, 8, cis-beta-carotene... 7, 8 trans-beta-carotene...7, 8, Carotenoids...7, 8, Cetyl betaine...,, Cetylpyridinium chloride...,,, Cetyltrimethylammonium chloride...,,, Chloramphenicol... 6 Chloride... Chlorohydrine... -Chloro--methylphenol... -Chlorophenol... Chlortetracycline... Cholecalciferol (Vitamin D)... 7, Chyrsene... Cinchonidine... 9 Citric acid...,, 7, Clonazepam... 9 Cohumulone... 9 Colupulone... 9 Cortisol (Hydrocortisone)..., Crotonaldehyde-DNPH... Curcumin... 7 Cytochrome-c tryptic digest... 6, 6 Cytosine... 8, Decyl sulfate...,,, Demethoxycurcumin Deoxycortisol... Dexamethasone... Dextromethorphan...,,-Diamino-6-nitrotoluene... 7 Diazepam... 9 Dibenzo[a,h]anthracene... Di-n-butylphthalate..., -Dichlorobenzidine..., 6,-Dichlorophenol... Diethyl heptadecyl imidazolinium ethylsulfate... Diethylphthalate... Dihydroquinine... 9,-Dihydroxybenzoic acid... 9,-Dihydroxybenzoic acid... 9,-Dihydroxybenzylamine... Dimethylaniline...7,,, -Dimethylbenzidine..., 6,-Dimethyl-,-dinitrobutane (DMDNB)... 7,-Dimethylphenol... Dimethylphthalate...,,-Dinitrobenzene...6, 7, 6,-Dinitrophenol...,-Dinitrophenylhydrazine (DNPH)...,-Dinitrotoluene...6, 7, 6,6-Dinitrotoluene...6, 7, 6 Di-n-octylphthalate... Diuron... Dodecyl gallate... Dodecyl sulfate...,,, Dodecylbenzene sulfonate..., Domiphen bromide... Dopamine... Doxepin..., 9, Doxycycline... Doxylamine..., Epicatechin gallate... 8 Epichatechin... 8 Epigallocatechin... 8 Epigallocatechin gallate... 8 Epinephrine... Epoxyefenone... Ergocalciferol (Vitamin D)... 7, Erythorbic acid... Ethoquad 8/... Ethyl -hydroxybenzoate... Ethyl benzoate... 7 Ethyl paraben... 9 Ethylbenzene... 7 Ethyleneglycol dinitrate (EGDN)... 7 Explosives...6, 7, 6 Fludrocortisone... Fluorene... Folic acid... 8, 9 Formaldehyde-DNPH... Formic acid... Formononotin... 8 Fumaric acid..., 7 Furaltadone... 8 Furazolidone... 8 Genistin... 8 Glutamate... Glutamine... 7 Gly-Tyr... 7 Heptanoic acid... Herbicides...,, 6 Hexahydroisohumulone... Hexaldehyde-DNPH... HMX...6, 7, 6 Humulone... 9 Hydrastine... 8 Hydrocortisone..., -Hydroxybenxoic acid... 9, -Hydroxycarbofuran..., 6 -Hydroxyethylbenzyl coco imidazolinium chloride... -Hydroxyindoleacetic acid... -Hydroxy--methoxymandelic acid... -Hydroxy--methoxyphenylacetic acid... Ibuprofen... Imipramine... 9, Indeno[,,-cd]pyrene... Insulin... 6 Ionox-... Isoadhumulone... Isobutyric acid... Isocaproic acid... Isocitric acid... Isocohumulone... Isohumulone... rho-isohumulone... Isoleucine... 7 Isovaleric acid... Ketoprofen... Labetalol..., 9, Lactic acid..., 7 Laureth sulfate...- Lauryl sulfate...,,, Lauryldimethylbenzyl ammonium chloride..., -, Laurylpyridinium chloride..., -, Leucine... 7 Leu-Enkephalin... 7 Lincomycin... 9 Linuron..., Loratidine... Lupulone... 9 trans-lutein... 8, - -

55 Analyte Index cis-lycopene... 8 nor-dehydroguiaretic acid (NDGA)... Propyl paraben..., 9 t-butylhydroquinone (TBHQ)... trans-lycopene... 7, 8 Norepinephrine... Protryptyline... 9 Temazepam... 9 Lysine... 7 Normetanephrine... Pseudo-ephedrine...,,, Terbutryn... 6 Lysozyme... 7 Nortriptyline... 9 Pyrene... o-terphenyl...7,, Maleic acid..., 9, ctyl gallate... Pyridine...7,, Tetracycline... Malic acid..., 7 ctyl methoxycinnamate... Pyridoxine... 8, 9 Tetrahydroisohumulone... Metanephrine... ctyl salicylate... Pyrrobutamine phosphate... 9 Tetryl...6, 7, 6 Metaraminol..., 9, Met-Enkephalin... 7 Methacrolein-DNPH... Methiocarb..., 6 Methomyl..., 6 -Methoxy--hydroxyphenylglycol... Methyl bis(ethyl(tallowate))--hydroxyethyl ammonium methylsulfate... -Methyl-,6-dichlorophenol... Methyl paraben..., 9 Methylethylketone-DNPH... Methylprednisolone... Metoprolol..., 9, Monuron..., Myoglobin... 6, 7 Myristamidopropylbetaine... N,N-Dimethylaniline...7,, Naphthalene..., Naproxen... NEDL Niacinamide... 8, 9 Nitrate..., Nitrobenzene...7,6, 7, 6 Nitrofurantoin... 8 Nitrofurazone Nitrophenol... -Nitrophenol... N-Nitrosodibutylamine... N-Nitrosodiethylamine... N-Nitrosodimethylamine... N-Nitrosodiphenylamine... N-Nitrosodipropylamine... N-Nitrosomethylethylamine... N-Nitrosomorpholine... N-Nitrosopiperidine... N-nitrosopyrrolidine... -Nitrotoluene...6, 7, 6 -Nitrotoluene...6, 7, 6 ctylphenoxyethoxyethyl dimethylbenzyl ammonium chloride...,,, il red... 9 nonin... 8 xalic acid... xamyl..., 6 xazepam... 9 xprenolol..., 9, xytetracycline... 6, Pantothenic acid... 8, 9 PEG monoethyl ether... PEG- Distearate... PEG-... PEG-... PEG-6... Pentachlorophenol... Pentaethyritol tetranitrate (PETN)... 7 Phenanthrene...,, Phenobarbital... Phenol...7,,, Phenols...7,,, Phenothiazine... 9 Phenytoin... PE() Lauryl sulfate... PE() Ammonium lauryl sulfate... Pramoxine... Prednisolone... Prednisone... Promethazine HCl... 9 Prometon... 6 Prometryn... 6 Propafenone... Propazine...,, 6 Propionaldehyde-DNPH... Propionic acid... Propoxur..., 6 Propranolol...,, 9,,, Propyl benzene... Quinine... 9 Ranitidine... Ranitidine S-oxide... RDX...6, 7, 6 trans-retinol... 7, Retinol acetate... 8, Riboflavin... 8, 9 Ribonuclease... 6 RNAse-A... 7 Saccharin Sodium... Secobarbital... Serotonin... Simazine... 6 Sissotrin... 8 Sodium lauryl sulfate...,,, Sorbic acid... 9 Spectinomycin... 9 Steroids..., Succinic acid...,, 7 Sudan range I... 9, 8 Sudan range II... 9, 8 Sudan range III... 9, 8 Sudan range IV... 9 Sulfachloropyridazine... Sulfadiazine..., Sulfadimethoxine...,, 6, Sulfamerazine...,, 6, Sulfamethazine...,, Sulfamethoxazole...,, Sulfanilamide..., Sulfanilic acid... Sulfapyridazine... Sulfapyridine... Sulfaquinoxaline..., Sulfathiazole..., Sulfisoxazole..., Tartaric acid... 7 Thenyldiamine HCl... 9 Theobromine... Thiamine... 8 Thymine...8,, 9 Timolol..., 9, alpha-tocopherol... 7, alpha-tocopherol acetate (Vitamin E)... 8, beta-tocopherol... 7 delta-tocopherol... 7 m-tolualdehyde-dnph... Toluene...7,,,,, Toluene sulfonate...,,6-trichlorophenol...,,-trihydroxybenzoic acid... 9 Trihydroxybenzophenone (THBP)...,,-Trinitrobenzene...6, 7, 6,,6-Trinitrotoluene...6, 7, 6 Triphenylene...7,, Triton X-...,,, Uracil..., 8,,,,,, Valeraldehyde-DNPH... Valeric acid... Valine... 7 Val-Tyr-Val... 7 Venlafaxine... 9 Vitamin A acetate... 8, Vitamin B... 8, 9 Vitamin C...7, 8, 9 Vitamin D... 7, Vitamin D... 7, Vitamin E... 7, Vitamin E acetate... 8, Vitamin K... Vitamins... 7 Xylene sulfonate...,, ZNYL FS fluorosurfactant... -Nitrotoluene...6, 7, 6 Propyl gallate... APPENDIX - -

56 Systems Acclaim Columns ptimal Performance with the UltiMate Intelligent LC and ICS- Corporate Headquarters Dionex Corporation 8 Titan Way P.. Box 6 Sunnyvale, CA Tel: (8) 77-7 Fax: (8) 7-9 The UltiMate Intelligent LC series delivers the full range of HPLC. Select from a wide range of modules to optimize systems for all types of applications, from sensitive nanoflow LC/MS analyses to automated semipreparative purifications. Whichever configuration you choose, you will get a highly integrated solution with optimum fluidic connections, single-point and intelligent control through Chromeleon software, and seamless intermodule communication. The UltiMate Intelligent LC System won: Gold award for best new product from Instrument Business utlook. The ICS- metal-free system is ideal for applications requiring chemically inert (PEEK) chromatography. The ICS- has won more than five awards: R&D Award as one of the best new products of 6 Two Gold Awards from Instrument Business utlook An IDEA Bronze award for Industrial design from Business Week's editorial staff. The Editors' Choice Silver Award as one of the best new products at Pittcon Worldwide Sales and Service North America U.S. (87) 9-7 Canada (9) 8-96 South America Brazil () 7 Europe Austria () 66 Belgium () 9 Denmark () France () 9 Germany (9) Italy (9) 6 67 The Netherlands () Switzerland () U.K. and Ireland () Asia Pacific Australia (6) 9 China (8) 8 8 India (9) 87 Japan (8) Korea (8) 6 8 Singapore (6) Dionex products are designed, developed, and manufactured under an IS 9 Quality System. 7 Dionex Corporation AQA and MSQ are trademarks of Thermo Electron Corporation. Zantac is a registered trademarks of Glaxo Smith Kline, Inc. All other trademarks and registered trademarks are the property of Dionex Corporation. LPN 668- PDF 7/7 Passion. Power. Productivity