YMC - Hichrom
Transcription
YMC - Hichrom
HICHROM Chromatography Columns and Supplies LC COLUMNS YMC Catalogue 9 Hichrom Limited 1 The Markham Centre, Station Road Theale, Reading, Berks, RG7 4PE, UK Tel: +44 (0)118 930 3660 Fax: +44 (0)118 932 3484 Email: sales@hichrom.co.uk www.hichrom.co.uk YMC LC Columns – YMC YMC YMC-Triart inorganic siloxane layer • Multi-layer hybrid material • Innovative micro-reactor technology • Multi-stage endcapping • High pH and temperature stability • 1.9, 3 and 5μm particle sizes YMC-Triart phases are based on a multi-layered hybrid material, manufactured using innovative micro-reactor technology. The combination of this layered hybrid particle with the precise microreactor granulation and the multi-stage endcapping procedures, produces a versatile material with outstanding chemical and physical durability, plus excellent chromatographic performance with acidic, basic and neutral compounds. organic siloxane layer Phase Specifications1 YMC-Triart C18 YMC-Triart C8 YMC-Triart Diol-HILIC Organic/inorganic silica Organic/inorganic silica Organic/inorganic silica C18 C8 Diol 1.9, 3, 5 1.9, 3, 5 1.9, 3, 5 120 120 120 Polymeric type Polymeric type - Multi-stage hybrid groups Multi-stage hybrid groups - C18: 16 (20% in total) C8: 7 (11% in total) - 1-12 1-12 2-10 pH 1-7:70°C, pH 7-12: 50°C pH 1-7:70°C, pH 7-12: 50°C 50°C Base Material Ligand Particle Size (μm) Pore Size (Å) Bonding Endcapping Carbon Load (%) pH Range Temperature Range 1 Other Triart phases are available. See page 6 Features of YMC-Triart • Homogeneous and uniform particles. Use of microreactor technology produces homogeneous and uniform particles that result in low operating pressure and reproducible surface modification, leading to excellent column-to-column and lot-to-lot reproducibility. • Multi-stage endcapping. YMC-Triart uses an innovative endcapping procedure for surface modification. Using a stepwise, multi-stage process with a number of compounds with different reactivities, the maximum number of silanols can be capped. • Balanced hydrophobicity. YMC-Triart exhibits a balanced hydrophobicity, making it more retentive than other hybrids. • Excellent durability. YMC-Triart C18 and C8 show excellent chemical stability. Even at high pH (up to pH 12) or high temperature conditions (up to 70°C), the column lifetime is greater than for conventional C18 and C8 columns. For free technical advice and support email technical@hichrom.co.uk • Method transfer between HPLC ↔ UHPLC. YMC-Triart phases provide matching chromatographic behaviour for all particle sizes. This ensures identical selectivity and excellent peak shape for basic compounds for all particle sizes. YMC-Triart allows predictable scale-up from UHPLC to conventional HPLC and even to semi-preparative LC and vice versa. Figure 1 illustrates method transfer from a 5μm, 150 x 2.0mm YMC-Triart C18 to a 1.9μm, 50 x 2.0mm column. Resolution is maintained but run time has been decreased from 55mins to 6mins. 266 A 1 4.5 MPa (650 psi) 5% B (0-3 min) 5-30% B (3-10 min) 30-45% B (10-55 min) 0.2 ml/min 2 0 3 10 7 4 20 55 min 8 9 5 6 30 40 Mins 55 50 1 B 2 37 MPa (5400 psi) 5% B (0-0.3 min) 5-30% B (0.3-1.0 min) 30-45% B (1.0-6.0 min) 0.6 ml/min 7 3 8 9 4 6 min 6 5 0 1 2 3 Figure 1. Method transfer between HPLC and UHPLC 4 5 6 Mins 1. Oxine copper 2. Asulam 3. Thiram 4. Triclopyr 5. Mecoprop 6. Flazasulfuron 7. Siduron 8. Halosulfuron-methyl 9. Azoxystrobin Columns:A: YMC-Triart C18 (5μm, 150 x 2.0mm) B: YMC-Triart C18 (1.9μm, 50 x 2.0mm) Eluent: A: 0.1% formic acid in H2O B: 0.1% formic acid in CH3CN Temperature: 40°C Detection: UV, 240nm YMC Features of YMC-Triart (continued) • Improvement of loadability. YMC-Triart C18 can tolerate larger injection volumes containing strong solvents (eg. acetonitrile), whilst maintaining better peak shape than traditional silica-based columns. This can be important for difficult to solubilise samples that need higher concentrations of organic solvent for solubilisation prior to analysis. Figure 2 illustrates the influence of injection volume on peak shapes. • LC-MS compatibility. Extremely low levels of column bleed lead to reduced background noise and hence lower levels of detection. In Figure 3, no bleed is observed in the test of total ion current measured by LC-MS with blank (no column) or with YMC-Triart C18. LC Columns – YMC YMC-Triart (continued) Solvent for sample: acetonitrile Injection volume: 1.0µl, 1.5µl, 2.0µl, 3.0µl Column: 5μm, 50 x 2.0mm Eluent: A: Water – formic acid (100:0.1) B: CH3CN – formic acid (100:0.1) 5 to 100% B from 0.5 to 2.5min Flow rate: 0.4ml/min Temperature: 40°C Detection: UV, 275nm Column: YMC-Triart C18 (5μm, 50 x 2.0mm) Eluent: A: 0.1% formic acid in H2O B: 0.1% formic acid in CH3CN Gradient: 5% B (0-1min), 5-100% B (1-5min), 100-5% B (10-10.1min), 5% B (10.1- 12.5min) Flow rate: 0.4ml/min Temperature: 40°C Detection: ESI +ve, TIC (mass range 50-1000) 1. Propranolol 2. Verapamil YMC-Triart C18 YMC-Triart C18 No column Silica-based C18 Figure 2. Influence of injection volume on peak shapes* Figure 3. LC-MS compatibility* *The comparative separations and data presented here may not be representative for all applications. Hardware for YMC-Triart 1.9μm columns YMC-Triart columns packed with 1.9μm particle size phases are provided in PT-hardware, which is compatible with Waters UPLC® systems and Agilent equipment. This enables optimised chromatographic separations to be achieved when using these and other instruments. Ordering Information When ordering please replace ‘XX’ with TA for Triart C18, TO for Triart C8 and TDH for Triart Diol-HILIC. 1.9μm YMC-Triart C18, C8 and Diol-HILIC Columns4 Column Length1 (mm) Column i.d. (mm) Guard Cartridges2 20 30 50 100 150 (5mm length) 2.0 XX12SP9-0202PT £469 XX12SP9-0302PT £454 XX12SP9-0502PT £469 XX12SP9-1002PT £483 XX12SP9-1502PT £528 XX12SP9-E5Q1CC £409 3.0 - XX12SP9-0303PT £469 XX12SP9-0503PT £469 XX12SP9-1003PT £483 - XX12SP9-E503CC £409 3µm and 5μm YMC-Triart C18, C8 and Diol-HILIC Columns4 Column i.d. (mm) Column Length1 (mm) 3μm Particle Size 50 150 250 (10mm length) 2.0 XX12S03-0502WT £320 XX12S03-1002WT £320 XX12S03-1502WT £379 XX12S03-2502WT £394 XX12S03-01Q1GC £231 3.0 XX12S03-0503WT £298 XX12S03-1003WT £342 XX12S03-1503WT £387 4.6 XX12S03-0546WT £305 XX12S03-1046WT £357 XX12S03-1546WT £417 XX12S03-2546WT £454 XX12S03-0104GC £231 5μm Particle Size 50 100 150 - XX12S03-0103GC £231 250 2.0 XX12S05-0502WT £312 XX12S05-1002WT £357 XX12S05-1502WT £372 XX12S05-2502WT £394 XX12S05-01Q1GC £231 3.0 XX12S05-0503WT £298 XX12S05-1003WT £335 XX12S05-1503WT £357 XX12S05-2503WT £402 XX12S05-0103GC £231 4.6 XX12S05-0546WT £305 XX12S05-1046WT £350 XX12S05-1546WT £379 XX12S05-2546WT £431 XX12S05-0104GC £231 3 4 Hichrom Limited Other dimensions available 2 3/pk. Requires holder XPCHUHP (£364) 1 100 Guard Cartridges3 5/pk. Requires holder XPGCH-Q1 (£132) - see p. 272 for further information YMC-Triart Phenyl and PFP also available - please enquire YMC-Triart preparative 10 and 15μm bulk material also available – please enquire. www.hichrom.co.uk • sales@hichrom.co.uk • Tel: 0118 930 3660 267 LC Columns – YMC YMC YMC ProFamily • Ultra high purity silica • C18, C8 and C4 bonded phases • Hydrosphere for aqueous stability • ProC18 and Hydrosphere C18 in 2, 3 and 5μm phases The ProFamily is YMC’s range of state-of-the-art ultra high purity silica phases (99.999%). They are manufactured using high specification silica with extremely low individual metal ion contents (typically <10ppm). Test certificates supplied for both the bulk material and the column characterise the quality and reproducibility of these products. YMC ProFamily Phases YMC Phase 1 Code Description Particle Size (μm) Pore Size (Å) Carbon Load (%) pH Range 21, 3, 5 120 16 2-8 Applications Fat soluble vitamins, antioxidants, acidic, neutral, basic and chelating compounds ProC18 AS Ultrapure silica base with low residual non-specific interactions ProC18RS RS High carbon load with polymeric bonding C18 3, 5 80 22 1-102 Hydrosphere C18 HS Can be used in 100% aqueous eluent 21, 3, 5 120 12 2-8 ProC8 OS Latest generation C8 with very low residual non-specific interactions 3, 5 120 10 2-7.5 Acidic, neutral, basic and chelating compounds, drugs and metabolites ProC4 BS Ultrapure silica base, endcapped 3, 5 120 7 2-7.5 Polar acidic, basic and chelating compounds, polar peptides YMC-UltraHT phases 2 Wide range of samples including acidic and basic compounds Strongly polar compounds, antibiotics, catecholamines, nucleic acids, water soluble vitamins It is recommended to use at least 10% organic solvent composition near the pH limits and over 50% at pH levels above pH 9.0 to preserve column lifetimes Phases in the YMC ProFamily are ideal for applications in both the pharmaceutical and biotechnology industries. ProC18RS is specifically designed for improved retention of non-polar, hydrophobic compounds. The high carbon load and surface area (510m2/g) and extended pH stability provide further selectivity options for demanding separations, including LC-MS and method development. The enhanced steric selectivity of this phase enables positional isomers to be separated. Hydrosphere C18 contains a hydrophilic C18 surface for enhanced polar selectivity and operation under 100% aqueous eluent conditions. Typical applications include nucleic acid bases, nucleotides, organic acids, catecholamines, vitamins etc. Figure 4. Comparison of YMC ProFamily separation selectivity For free technical advice and support email technical@hichrom.co.uk Figure 4 compares the selectivity characteristics of the three C18 phases using hydrophobicity and hydrogen-bonding ability as indicators. This plot highlights the contrasting separation features of these phases. Figure 5 shows a comparison of the differing hydrophobicities and hydrogen bonding capacities of each phase in combination with basic substances. 268 1 Column: 50 x 2.0mm Eluent: CH3OH - 10mM CH3COONH4 (55:45) Flow rate: 0.2ml/min Temperature: 37˚C Detection: UV at 254nm ProC4 3 2 1 ProC8 4 1 2 5,4 Hydrosphere C18 1 ProC18 2 ProC18RS 2 3 5 1. Toluene 2. Amitriptyline HCl 3. Progesterone 4. Propylbenzene 5. Acenaphthene 1 3 3 2 3 4 4 5 4 5 5 0 10 Mins lowest hydrophobicity 0 10 20 lower hydrophobicity Mins 0 10 20 balanced hydrogen bonding, capacity and hydrophobicity 30 Mins 0 10 20 30 highest carbon load Figure 5. Comparison of characteristics of ProFamily phases 40 Mins 0 10 20 Mins high polar recognition YMC • ProC18 and Hydrosphere C18 phases • Same selectivity as the corresponding 3 and 5μm phases • Use with UHPLC or conventional HPLC systems • Fully downscalable from 3 and 5μm to 2μm The YMC-UltraHT ProC18 and Hydrosphere C18 bonded phases are based on high efficiency 2μm spherical silica particles, of the same pore size, surface area and carbon content as the equivalent 3 and 5μm products (see page 268). They are specifically designed for high speed and high throughput analyses in various fields including pharmaceuticals, foods and environmentals. YMC-UltraHT columns exhibit low pressure drop and high flow rates can be achieved without generating excessive back pressure and without the need for specialised equipment. LC Columns – YMC YMC-UltraHT (2μm) Due to the production process used to manufacture the YMC ProFamily, methods can easily be downscaled without changes to selectivity. Figure 6 shows the transfer of a separation of a polar isoflavone mixture from a 5μm, 150 x 4.6mm i.d. YMC Hydrosphere C18 column to a 2μm, 50 x 2.0mm i.d. YMC-UltraHT Hydrosphere C18 column. A. Conventional LC method YMC Hydrosphere C18 5µm B. Ultra fast LC method YMC-UltraHT Hydrosphere C18 2µm Column: YMC Hydrosphere C18 (5µm, 150 x 4.6mm) Eluent: A: H2O - CH3CO2H (100:3) B: CH3CN - CH3CO2H (100:3) Gradient: 15 - 35% B in 30 mins Flow rate: 1.0ml/min Temperature: 35°C 3 9 1 2 0 6 4 5 6 1. Daidzin 2. Glycitin 3. Genistin 4. 6’-O-Malonyldaidzin 5. 6’-O-Malonylglycitin 6. 6’-O-Acetyldaidzin 7. 6’-O-Acetylglycitin 8. 6’-O-Malonylgenistin 9. Daidzein 10. Glycitein 11. 6’-O-Acetylgenistin 12. Genistein 12 10 11 78 12 18 24 Mins Column: YMC-UltraHT Hydrosphere C18 (2µm, 50 x 2.0mm) Eluent: A: H2O - CH3CO2H (100:3) B: CH3CN - CH3CO2H (100:3) Gradient: 12.5 - 30% B in 3.3 mins. Flow rate: 0.9ml/min Temperature: 35°C 1 3 9 2 4 0 5 1 12 10 11 6 7 8 2 3 Mins Figure 6. Downscaling of isoflavone separation Ordering Information – YMC ProFamily YMC Phase 3μm 50 x 2.1 AS12S03-05Q1QT ProC18 £335 RS08S03-05Q1QT ProC18RS £312 HS12S03-05Q1QT Hydrosphere C18 £312 OS12S03-05Q1QT ProC8 £312 BS12S03-05Q1QT ProC4 £312 5μm 50 x 2.1 AS12S05-05Q1QT ProC18 £327 RS08S05-05Q1QT ProC18RS £312 HS12S05-05Q1QT Hydrosphere C18 £312 OS12S05-05Q1QT ProC8 £312 BS12S05-05Q1QT ProC4 £312 YMC-UltraHT Phase 2μm ProC18 Hydrosphere C18 1 100 x 2.1 AS12S03-10Q1QT £335 RS08S03-10Q1QT £335 HS12S03-10Q1QT £335 OS12S03-10Q1QT £335 BS12S03-10Q1QT £335 150 x 2.1 AS12S05-15Q1QT £387 RS08S05-15Q1QT £387 HS12S05-15Q1QT £387 OS12S05-15Q1QT £387 BS12S05-15Q1QT £387 30 x 2.0 £537 AS12S02-0302WT HS12S02-0302WT 30 x 3.0 - 150 x 2.1 AS12S03-15Q1QT £402 RS08S03-15Q1QT £402 HS12S03-15Q1QT £402 OS12S03-15Q1QT £402 BS12S03-15Q1QT £402 250 x 2.1 AS12S05-25Q1QT £431 RS08S05-25Q1QT £409 HS12S05-25Q1QT £409 OS12S05-25Q1QT £409 BS12S05-25Q1QT £409 50 x 2.0 £512 AS12S02-0502WT HS12S02-0502WT (£521) 50 x 3.0 £521 AS12S02-0503WT HS12S02-0503WT (5/pk, 10 x 4mm) 50 x 4.6 AS12S03-0546WT £320 RS08S03-0546WT £312 HS12S03-0546WT £320 OS12S03-0546WT £312 BS12S03-0546WT £312 50 x 4.6 AS12S05-0546WT £320 RS08S05-0546WT £312 HS12S05-0546WT £320 OS12S05-0546WT £312 BS12S05-0546WT £312 100 x 4.6 AS12S03-1046WT £305 RS08S03-1046WT £357 HS12S03-1046WT £372 OS12S03-1046WT £357 BS12S03-1046WT £327 150 x 4.6 AS12S05-1546WT £387 RS08S05-1546WT £379 HS12S05-1546WT £402 OS12S05-1546WT £379 BS12S05-1546WT £379 Column Dimensions (mm) 75 x 2.0 £529 AS12S02-L502WT HS12S02-L502WT 75 x 3.0 £529 AS12S02-L503WT HS12S02-L503WT Use with universal holder XPGCH-Q1 (£132) or direct connect holder XPGCS-Q1 (£132) – see p. 272. 2.1mm i.d. guard cartridges also available 150 x 4.6 AS12S03-1546WT £446 RS08S03-1546WT £431 HS12S03-1546WT £446 OS12S03-1546WT £431 BS12S03-1546WT £431 250 x 4.6 AS12S05-2546WT £454 RS08S05-2546WT £454 HS12S05-2546WT £454 OS12S05-2546WT £454 BS12S05-2546WT £454 100 x 2.0 £537 AS12S02-1002WT HS12S02-1002WT 100 x 3.0 £537 AS12S02-1003WT HS12S02-1003WT 2 AS12S03-0104GC £245 RS08S03-0104GC £245 HS12S03-0104GC £245 OS12S03-0104GC £245 BS12S03-0104GC £245 AS12S05-0104GC £245 RS08S05-0104GC £245 HS12S05-0104GC £245 OS12S05-0104GC £245 BS12S05-0104GC £245 150 x 2.0 £678 AS12S02-1502WT HS12S02-1502WT 150 x 3.0 £711 AS12S02-1503WT HS12S02-1503WT Other dimensions (eg. 1.0mm i.d.) also available www.hichrom.co.uk • sales@hichrom.co.uk • Tel: 0118 930 3660 Hichrom Limited ProC18 Hydrosphere C18 Guard Cartridges1 Column Dimensions2 (mm) 269 For free technical advice and support email technical@hichrom.co.uk LC Columns – YMC YMC 270 YMC ‘Classical’ Phases YMC manufacture a wide range of ‘classical’ and speciality bonded phases for reversed-phase, normal-phase and HILIC methods. The traditional ODS-A, ODS-AL, ODS-AM and ODS-AQ phases are well documented in the scientific literature. The series of three J’sphere phases offers alternative hydrophobicities due to variation in degree of ODS substitution. The Polyamine ll material is a mixed secondary and tertiary amino derivative, covalently bonded onto 5μm particle size silica. This phase offers increased lifetime and alternative selectivity for carbohydrate applications compared with conventional primary amino materials. The PVA-SIL material is prepared by bonding the silica support with a monomolecular polymer coating of vinyl alcohol. It offers an excellent alternative selectivity to silica and can be used over a wider pH range. 1 Particle1 Size (μm) Pore Size (Å) Carbon Load (%) pH Range C30 ligands. Proprietary polymeric bonding chemistry 3, 5 proprietary - 2-6 AA Traditional high performance C18 3, 5 120, 200, 300 17, 12, 7 2-7.5 Pharmaceuticals, vitamins, amino acids, peptides, general purpose phase AM Strict QC controlled production provides high performance C18 column for validated methods 3, 5 120 17 2-7.5 Purines, phenols, PTC-amino acids, angiotensins, alkaloids ODS-AQ AQ ‘Hydrophilic’ endcapping for 100% aqueous eluents, increased retention of polar compounds 3, 5 120, 200 14, 11 2-7.5 Strong polar compounds, pharmaceuticals, antibiotics, peptides and proteins, nucleic acids ODS-AL AL Traditional C18 for ‘mixed-mode’ separations 3, 5 120 17 2-7.5 Tocopherols, fat soluble vitamins, disinfectants J’sphere JH, JM, JL C18 family with different hydrophobicity 4 80 22, 14, 9 (JH, JM, JL) 6 proprietary C18=10% 2-13 Phenols, anilines, peptides in high pH, pharmaceuticals, quaternary amines YMC Phase Code Carotenoid CT ODS-A ODS-AM Description Applications Isomeric carotenes, retinols, steroids, fat-soluble vitamins 1-9 (JH), Positional isomers, complexing 2-7.5 (JM, JL) agents, pharmaceuticals Polymer C18 PC Polymethacrylate matrix, stable towards shrinking and swelling, wide pH applicability C8 OC High coverage monomeric C8 bonding 3, 5 120, 200, 300 10, 7, 4 2-7.5 Proteins and peptides, estrogens, general purpose phase YMCbasic BA Specifically designed for separation of basic compounds 3, 5 proprietary 8 2-7.5 Basic molecules without modifiers, anilines, alkaloids, antidepressants Phenyl PH Monomeric phenyl bonding, endcapped 3, 5 120, 300 9, 3 2-7.5 Phenols, fullerenes, sweeteners C4 BU C4, fully endcapped, high coverage monomeric bonding 3, 5 120, 200, 300 7, 5, 3 2-7.5 Biological separations, polar compounds Protein RP PR Designed to withstand exposure to TFA, good recovery rates 5 proprietary 4 1.5-7.5 C1 TM Trimethylsilane, excellent hydrolytic stability 3, 5 120, 300 4, 3 2-7.5 PVA-SIL PV Polyvinyl alcohol bonded on silica support, suitable for NP or RP applications 5 120 - 2-9 Proteins, phospholipids, retinoids, lipids Polyamine ll PB Improved hydrolytic stability over conventional amino phase, increased lifetime, rapid equilibration time 5 120 - 2-9 Maltooligosaccharides, tocopherols, nucleotides, sugars Amino NH Primary amino, high coverage monomeric bonding chemistry 3, 5 120 3 2-7.5 Saccharides, nucleotides, water soluble vitamins Cyano CN Fully endcapped, also useful for SFC applications 3, 5 120, 300 7, 2.5 2-7.5 Proteins, steroids, catechols Diol DL For aqueous GPC or normalphase applications, high recovery for biological material 5 60, 120, 200, 300 9, 7, 5, 3 2-7.5 Peptides, proteins, maltooligosaccharides Silica SL Ultra high purity, high mechanical stability 3, 5 60, 120, 200, 300 - 2-7.5 Small organic molecules, fat soluble vitamins, tocopherols See page 274 for YMC preparative grade materials Proteins, peptides Water soluble vitamins YMC YMC C18 ODS-A This phase is the traditional workhorse for most reversed-phase applications. Endcapping is exhaustive and this greatly reduces the activity of silanol groups and minimises non-specific secondary interactions. YMC C18 ODS-AM YMC C18 ODS-AM is a high specification C18 material produced to meet the stringent demands for validated analytical HPLC processes. All columns have to pass additional tests to meet the narrow quality specifications range with regards to selectivity pattern, column resolution, absolute retention and peak symmetry. LC Columns – YMC YMC ‘Classical’ Phases (continued) YMC C18 ODS-AL This phase is a non-endcapped ODS. Non-endcapping imparts some additional secondary normal-phase adsorption interaction characteristics. YMC ODS-AQ ODS-AQ differs from traditional C18 phases in that after initial octadecylsilyl bonding, the material is treated to produce a surface that can be wetted with polar eluents. In highly aqueous eluents ODS-AQ gives longer retention times for polar compounds than conventionally endcapped ODS packings. YMC Polymer C18 Polymer C18 is a reversed-phase material which provides a broad range of solvent choices over a pH range of 2 – 13, due to its resistance to shrinking and swelling. It consists of a highly cross-linked spherical hydrophilic methacrylate polymer which is functionalised with C18 groups. Polymer C18 provides similar, but not identical, selectivity to standard silica-based ODS materials. It has slightly greater retention for aromatic compounds than even highly retentive ODS columns. It is significantly less retentive than styrene/divinylbenzene based HPLC columns. YMCbasic YMCbasic is an early base deactivated phase, widely used for the analysis of pharmaceutical and other basic compounds. The phase consists of monomerically bonded alkyl chains with different chain lengths of C8 and smaller, leading to high surface coverage. Its selectivity is similar to that expected for conventionally endcapped C8 reversed-phase media. YMC J’sphere In the YMC J’sphere series of phases, instead of varying the alkyl group chain length in order to produce phases of different polarity, the surface coverage of C18 is varied to produce three phases of graduated hydrophobicity and hydrogen bonding characteristics. The stepwise decrease of hydrophobicity in the J’sphere ODS-H80, M80 and L80 series is accompanied by a corresponding increase in the H-bonding capacity. YMC Carotenoid The YMC Carotenoid (C30) phase is very lipophilic with exceptional selectivity towards isomeric structures. The C30 ligands provide sufficient phase thickness to enhance interaction with long chained molecules. Geometrical and positional isomers of conjugated double bond systems can be recognised and resolved. The phase is particularly suited for the separation of carotenoids (see Figure 7). Other potential applications include the separation of prostaglandins and leukotrienes. 1. Lutein 2. Zeaxanthin 3. ß-Cryptoxanthin 4. 13-cis ß-Carotene 5. α-Carotene 6. trans ß-Carotene 7. 9-cis ß-Carotene 8. Lycopene Column: YMC Carotenoid (5µm, 250 x 4.6mm) Eluent: A: CH3OH - MTBE - H2O (81:15:4) B: CH3OH - MTBE - H2O (6:90:4) Gradient A to B in 90mins Flow rate: 1ml/min Temperature: Ambient Detection: UV, 450nm 8 6 1 3 2 5 4 0 10 20 30 40 7 50 60 70 80 Mins Figure 7. Extract of SRM 2383, NIST food standard Hichrom Limited www.hichrom.co.uk • sales@hichrom.co.uk • Tel: 0118 930 3660 271 LC Columns – YMC YMC Ordering Information - YMC ‘Classical’ 120Å Phases YMC2 Phase 3μm Particle Size 50 x 3.0 100 x 3.0 150 x 3.0 100 x 4.6 150 x 4.6 - ODS-AQ ODS-A AA12S03-0503QT AA12S03-1003QT AA12S03-1503QT AA12S03-0546WT AA12S03-1046WT AA12S03-1546WT AA12S03-0104GC £314 £347 £388 £347 £380 £405 £273 4μm Particle Size 50 x 3.0 150 x 3.0 250 x 3.0 J’sphere ODS-L805 JL08S04-0503QT £355 JL08S04-1503QT £455 JL08S04-2503QT £537 J’sphere ODS-M805 JM08S04-0503QT JM08S04-1503QT JM08S04-2503QT JM08S04-0546WT JM08S04-1546WT JM08S04-2546WT JM08S04-0104GC £355 £455 £537 £372 £496 £521 £273 J’sphere ODS-H805 JH08S04-0503QT £355 JH08S04-1503QT £455 50 x 4.6 150 x 4.6 250 x 4.6 - 50 x 3.0 150 x 3.0 JL08S04-0546WT JL08S04-1546WT JL08S04-2546WT £372 £496 £521 JL08S04-0104GC £273 JH08S04-2503QT JH08S04-0546WT JH08S04-1546WT JH08S04-2546WT JH08S04-0104GC £537 £372 £496 £521 £273 250 x 3.0 50 x 4.6 150 x 4.6 250 x 4.6 - ODS-AQ AQ12S05-0503QT AQ12S05-1503QT AQ12S05-2503QT AQ12S05-0546WT AQ12S05-1546WT AQ12S05-2546WT AQ12S05-0104GC £314 £380 £413 £339 £405 £430 £273 ODS-A AA12S05-0503QT AA12S05-1503QT AA12S05-2503QT AA12S05-0546WT AA12S05-1546WT AA12S05-2546WT AA12S05-0104GC £314 £380 £405 £306 £372 £405 £273 ODS-AM AM12S05-0503QT AM12S05-1503QT AM12S05-2503QT AM12S05-0546WT AM12S05-1546WT AM12S05-2546WT AM12S05-0104GC £314 £380 £413 £339 £405 £430 £273 ODS-AL AL12S05-0503QT £339 AL12S05-1503QT £380 50 x 3.0 150 x 3.0 6μm Particle Size Polymer C18 AL12S05-2503QT AL12S05-0546WT AL12S05-1546WT AL12S05-2546WT AL12S05-0104GC £430 £339 £413 £446 £273 250 x 3.0 50 x 4.6 150 x 4.6 250 x 4.6 - PC99S06-0503QT PC99S06-1503QT PC99S06-2503QT PC99S06-0546WT PC99S06-1546WT PC99S06-2546WT PC99S06-0104GC £372 £496 £537 £413 £545 £521 £273 Column Dimensions4 (mm) YMC2, 3 5μm Phase 100 x 4.6 Guard Cartridges1 150 x 4.6 250 x 4.6 (5/pk, 10 x 4mm) Carotenoid (C30) CT99S05-1046QT £413 CT99S05-1546QT £496 CT99S05-2546QT £554 CT99S05-0104GC £273 C8 OC12S05-1046WT £388 OC12S05-1546WT £413 OC12S05-2546WT £446 OC12S05-0104GC £273 Basic BA99S05-1046WT £413 BA99S05-1546WT £446 BA99S05-2546WT £504 BA99S05-0104GC £273 C4 BU12S05-1046WT £388 BU12S05-1546WT £413 BU12S05-2546WT £446 BU12S05-0104GC £273 TMS TM12S05-1046WT £405 TM12S05-1546WT £405 TM12S05-2546WT £446 TM12S05-0104GC £273 Phenyl PH12S05-1046WT £405 PH12S05-1546WT £405 PH12S05-2546WT £446 PH12S05-0104GC £273 Protein RP PR99S05-1046WT £521 PR99S05-1546WT £521 PR99S05-2546WT £562 PR99S05-0104GC £273 SIL SL12S05-1046WT £347 SL12S05-1546WT £372 SL12S05-2546WT £405 SL12S05-0104GC £273 PVA-SIL PV12S05-1046WT £504 PV12S05-1546WT £479 PV12S05-2546WT £554 PV12S05-0104GC £273 Polyamine ll PB12S05-1046WT £413 PB12S05-1546WT £446 PB12S05-2546WT £504 PB12S05-0104GC £273 NH2 NH12S05-1046WT £405 NH12S05-1546WT £405 NH12S05-2546WT £446 NH12S05-0104GC £273 CN CN12S05-1046WT £405 CN12S05-1546WT £405 CN12S05-2546WT £446 CN12S05-0104GC £273 Diol - - DL12S05-2546WT £504 - Use with universal holder XPGCH-Q1 (£132) or direct connect holder XPGCS-Q1 (£132). 3mm i.d. guard cartridge also available 2 60Å, 200Å or 300Å pore sizes also available for selected materials 3 Available in 3μm particle size for selected materials 1 For free technical advice and support email technical@hichrom.co.uk (5/pk, 10 x 4mm) 50 x 4.6 AQ12S03-0503QT AQ12S03-1003QT AQ12S03-1503QT AQ12S03-0546WT AQ12S03-1046WT AQ12S03-1546WT AQ12S03-0104GC £339 £355 £405 £339 £405 £413 £273 5μm Particle Size 272 Guard Cartridges1 Column Dimensions4 (mm) Other dimensions (eg 2.1 and 1.0mm i.d.) also available 5 80Å 4 YMC Guard Cartridge Holders For 4.6mm i.d. analytical columns, the universal holder XPGCH-Q1 is required for 10mm length guard cartridges and XPGCH-Q2 for 20mm length guard cartridges (see figure 8). For 1.0 to 4.0mm i.d. guard cartridges, the customer can choose between this universal holder or an integral (direct connect) holder XPGCS-Q1 for 10mm length cartridges and XPGCS-Q2 for 20mm length guard cartridges (see figure 9). The integral holder replaces the inlet endfitting on the column with a hand-tightened holder which allows for a very low dead volume connection. Figure 8. Universal holder XPGCH-Q1 or XPGCH-Q2 Figure 9. Integral holder XPGCS-Q1 or XPGCS-Q2 YMC LC Columns – YMC YMC-BioPro • Porous and non-porous hydrophilic polymer • Ion-exchange phases • Excellent binding capacity and recovery • High resolution YMC-BioPro is a series of porous and non-porous ion-exchange phases based on a hydrophilic polymer with low non-specific adsorption. The phases are designed for the analysis and purification of peptides, proteins, nucleic acids and other biomolecules. They also show higher binding capacity and higher recovery of biomolecules compared to conventional ion-exchange columns. The completely spherical, monodisperse beads, together with optimal packing technology, provide high efficiency and symmetrical peak shapes. YMC-BioPro Phases YMC Phase QA SP QA-F SP-F Pore Size (Å) 1000 1000 Non-porous Non-porous 5 5 5 5 -CH2N (CH3)3 Cl- -CH2CH2CH2SO3 -CH2N (CH3)3 Cl- -CH2CH2CH2SO3 Particle Size (μm) Charged Group + Counter Ion pH Range Dynamic Binding Capacity + Na+ Na+ 2-12 2-12 2-12 2-12 110-150mg BSA/ml resin 70-100mg human IgG/ml resin 12-20mg BSA/ml resin 10-18mg human IgG/ml resin The porous YMC-BioPro phases show high dynamic binding capacity and excellent recovery, making them useful for semi-preparative separations of proteins and antibodies. Figure 10 shows a loading study for proteins on a YMC-BioPro QA column. Figure 11 shows the high resolution separation of DNA fragments achievable with a YMC-BioPro QA-F column. Column:YMC-BioPro QA (5μm, 50 x 4.6mm) Eluent: A. 20mM Tris-HCl (pH 8.1) B. 20mM Tris-HCl (pH 8.1) containing 0.5M NaCl 10-80% B over 30 minutes Flow rate: 0.5ml/min 1 Temperature: 25°C Detection: UV, 280nm 1. Ovalbumin 2. Trypsin inhibitor (soybean) Column: YMC-BioPro QA-F (5μm, 100 x 4.6mm) Eluent: A. 20mM Tris-HCl (pH 8.1) containing 0.5M NaCl B. 20mM Tris-HCl (pH 8.1) containing 1.0M NaCl 40-100% B over 30 minutes Flow rate: 0.5ml/min Temperature: 25°C Detection: UV, 260nm 1kb DNA ladder (75-12,216 bp) 2 500µg 250µg 100µg 50µg 0 10 20 Mins 30 0 5 10 15 Mins 20 Figure 11. DNA fragments analysed on YMC-BioPro QA-F Figure 10. Loading study for YMC-BioPro QA Ordering Information1 Column Dimensions (mm) SP QA-F SP-F 30 x 4.6 - - QF00S05-0346WP £760 SF00S05-0346WP £760 50 x 4.6 QAA0S05-0546WP £926 SPA0S05-0546WP £926 £893 SF00S05-0546WP £893 100 x 4.6 QAA0S05-1046WP £992 - QF00S05-0546WP QF00S05-1046WP £1,017 SF00S05-1046WP £1,017 Preparative grade YMC-BioPro also available as bulk material www.hichrom.co.uk • sales@hichrom.co.uk • Tel: 0118 930 3660 Hichrom Limited 1 YMC-BioPro Phase QA 273 LC Columns – YMC YMC YMC Capillary Columns YMC manufacture a range of LC-MS compatible nano and capillary columns of 75, 100, 300 and 500μm i.d. and lengths of 50, 75, 100 and 150mm. For highly contaminated samples, the use of pre-columns is recommended. These connect to the capillary column with a pre-column coupler which is included. Pre-columns can also be used as trapping columns. Figure 12. Capillary column with pre-column attached YMC Chiral Columns YMC manufacture a range of chiral columns, based on different ligand types, suitable for a wide range of chiral separations. See also page 5 for the new YMC CHIRAL polysaccharide series of columns. YMC Chiral Phases YMC Phase Ligand Particle Size (μm) Pore Size (Å) Recommended pH Range 1-Naphthylethylamine 5 300 2 - 6.5 (RP) Bromide derivative of cyclodextrin (α, β or γ) 5 120 3.5 - 6.5 Various – 17 different types 5 120 - Chiral NEA (R) or (S) CD BR (α, β or γ) Sumichiral OA Series YMC Chiral NEA (R) and (S) YMC Chiral NEA consists of 1-naphthylethylamine bonded to wide pore spherical silica. (R) and (S) versions are available for elution order reversal. YMC Chiral NEA columns can be used in normal-phase and reversed-phase modes. YMC Chiral CD BR YMC Chiral CD BR phases consist of bromide derivatives of α-, β- or γ-cyclodextrin, in which the primary hydroxyl groups at carbon 6 are substituted for bromine. These phases are used in reversed-phase mode to separate a wide range of polar, water-soluble compounds and positional isomers of substituted aromatic compounds. YMC Sumichiral OA Seventeen different phases of Sumichiral OA columns are available. These are based on different types of covalent linkage, including amide, urea, ligand exchange type, cyclodextrin type and crown ether phases. Please contact Hichrom for ordering information for YMC chiral columns. YMC Chiral b-CD BR For free technical advice and support email technical@hichrom.co.uk YMC SFC Columns 274 In addition to its application in enantiomer purification, SFC is gaining importance in the separation of achiral compounds. YMC offer six different selectivity phases for achiral SFC: • Diol • Amino • Cyano • PVA-Sil • Polyamine ll • Silica For preparative SFC, YMC has developed special column hardware in order to meet the highest demands in quality, safety and reliability. Please contact Hichrom for further information on YMC SFC columns. YMC Preparative Separations YMC preparative columns, packed with a variety of YMC materials, are available in a wide range of column sizes to accommodate virtually any preparative separation and direct scale-up from analytical and semi-preparative separations. Standard column dimensions range from 50 to 200mm i.d. and 250 to 1000mm in length. Matching YMC preparative guard columns are available. Please contact Hichrom for further information. YMC • Purification and isolation of maximum sample quantity • Savings in time and costs • Excellent durability and efficiency YMC-Actus semi-preparative/preparative columns use modified hardware and packing technology. These columns are specifically designed for fast semi-preparative/preparative chromatography and are available in a range of dimensions from 50 x 20mm i.d. to 250 x 50mm i.d. A uniform packing density results in a longer lifetime than conventional semi-preparative/preparative columns. YMC-Actus columns provide reliable results, even after exposure to severe, rapid gradient conditions and multiple injections. LC Columns – YMC YMC-Actus for Fast Semi-preparative/Preparative Separations Figure 13 illustrates the excellent durability and efficiency of the purification of glycyrrhizic acid from a herbal medicine achieved under fast gradient conditions at high flow rate. Analysis YMC-Pack ODS-AQ, 50 x 4.6 mm i.d., 5µm 3ml/min, 10µl injection Glycyrrhizic acid Glycyrrhizic acid 0 5 10 Mins Purification YMC-Actus ODS-AQ, 50 x 20 mm i.d., 5µm 60ml/min, 500µl injection After purification Glycyrrhizic acid 99.5% 0 0 5 5 10 10 Mins Mins Eluent: A: Water – acetic acid (99:1) B: Methanol – acetic acid (99:1) Gradient: 20% B (0 – 2mins), 20-45% B (2 – 7 mins), 45% B (7 – 10mins) Temperature: Ambient Detection: UV, 260nm Sample: Water – methanol – acetic acid extract of commercially available herbal medicine (0.1g/ml) Figure 13. Purification of glycyrrhizic acid from herbal medicine Ordering Information1 (5μm, 50 x 20mm i.d. Columns) Phase 1 Pore Size (Å) Part Number Price Phase Pore Size (Å) Part Number Price Triart C18 120 TA12S05-0520WX £959 ProC18RS 80 RS08S05-0520WX £959 Triart C8 120 TO12S05-0520WX £959 ODS-A 120 AA12S05-0520WX £959 ProC18 120 AS12S05-0520WX £959 ODS-AQ 120 AQ12S05-0520WX £901 Hydrosphere C18 120 HS12S05-0520WX £959 ProC8 120 OS12S05-0520WX £959 Other phases and dimensions available. Please contact us for details. Bulk Material YMC supply a wide range of bulk media for scale-up to preparative dimensions and beyond. These include materials for reversed-phase, normal-phase, SEC, ion-exchange and chiral. A selection of media is listed below. Please contact us for media not listed. HG stands for ‘High Grade’ 1 Particle Size (μm) 10, 15, 20, 50 10 10, 15, 20, 50 10, 15, 20 10, 30, 75 10, 20, 50 10 Pore Size1 (Å) 120, 200 120 80, 120, 200, 300 120, 200, 300 1000 120 300 Not all combinations of particle size and pore size are available www.hichrom.co.uk • sales@hichrom.co.uk • Tel: 0118 930 3660 Hichrom Limited Material Triart Prep C18, C8 ProC18 ODS-A-HG, ODS-AQ-HG, C8-HG, C4-HG, NH2-HG, CN-HG, Diol-HG, SIL-HG, Ph-HG, TMS-HG Diol BioPro QA, SP Chiral Prep CD ST and CD PM Chiral Prep NEA 275
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