WA30000 HPLC Columns Applications Notebook Table of Contents Click on the items below to go directly to that section of the notebook. To Start a new search, click on Table of Contents Bookmark to the left YMC™ HPLC Column Quality Assurance............................................................2 Guide to YMC™ Part Numbering System ...........................................................3 YMC™ Column Selection Guide ....................................................................4-7 HPLC Troubleshooting and Reference Guide..................................................8-14 List of Applications..................................................................................15-20 1. Vitamins .....................................................................................21 2. Carbohydrates ............................................................................32 3. Nucleic Acids .............................................................................43 4. Amino Acids, Peptides, Proteins......................................................48 5. Organic Acids, Fatty Acid Derivatives .............................................60 6. Food Additives ............................................................................62 7. Natural Products ..........................................................................68 8. Steroids......................................................................................77 9. Drugs, Metabolites .......................................................................87 10. Antibiotics.................................................................................103 11. Pesticides, Environmental .............................................................114 12. Miscellaneous ...........................................................................119 Index of Analytes.......................................................................................128 1 YMC™ HPLC Column Quality Assurance Waters Establishes Worldwide Distribution Network for YMC™ HPLC Columns Waters Corporation announces worldwide distribution of the YMC™ HPLC A Product You Can Count On analytical and preparative column lines, a highly respected series of prod- ™ ucts that Waters has distributed since 1997. In expanding the distribution YMC products must pass internal performance criteria as raw materials, of YMC™ columns globally, Waters maintains exclusive distribution rights as synthesized silica particles, after they are bonded, and finally when for the YMC™ column product line in the Americas (North America, Central they are packed into a column. By setting tight standards, testing the raw America, South America and the Caribbean Islands). materials and precise control of silica gel synthesis, low metal levels of puri- ty are achieved. Acceptance standards virtually eliminate problems with YMC...a Premium HPLC Packing Manufacturer nonspecific adsorption caused by trace contaminants in the silica gel. YMC has built its reputation on the supply of consistent, premium quality HPLC packing and columns. Control of the silica support is essential for product consistency and is of the highest priority to YMC. The silica sup- ports are then functionalized with the Company’s proprietary bonding and endcapping chemistries to yield rugged, reproducible, high performance HPLC packings. Consistent Production Methods Year to Year Once a YMC product has been provided to customers for their chro- matography, the methods used to produce that product will not be changed, as a matter of company policy. Even small changes in manu- facturing methods to “improve” a product can lead to selectivity changes in methods for which that column is applied. Because YMC controls the pro- duction of its products from silica synthesis to packed column you can be assured that a method developed on a YMC™ column will be reproducible for as long as you conduct that method. In-house manufacture gives YMC complete control over product quality and reproducibility. Proven procedures at every stage of manufacture guaran- tee consistency in silica purity, particle size, pore size, bonding and end- capping. Test chromatograms shipped with every column are merely the YMC-Pack™ Pro C18™ columns come with a detailed Certificate of final demonstration of a rigorously controlled product. Analysis for reliability and traceability. US Pharmacopeia HPLC "L1" (C18) Column Designation Subclassification Testing by Brand Name* Hydrophobicity Chelating Silanol Activity Shape Selectivity Column Brand Name (k, Ethylbenzene) (Tailing Factor (TF), Quinizarin) (k, Amitriptyline) (TF, Amitriptyline) (Bonding Density, mmol/m2) Pro C18™ 1.88 1.28 4.74 1.50 2.50 ™ Hydrosphere C18 1.52 1.34 4.40 1.94 1.90 Pro C18™ RS 3.28 1.17 7.50 1.35 3.04 ODS-AL™ 1.79 1.52 13.59 2.81 2.56 ODS-A™ 1.89 1.53 5.00 2.40 2.66 ODS-AM™ 1.94 1.52 5.11 2.40 2.75 ODS-AQ™ 1.88 1.49 5.96 2.86 2.22 J’sphere™ ODS-H80 3.32 1.31 8.80 2.93 3.18 J'sphere™ ODS-M80 1.62 1.41 4.96 2.72 1.02 J’sphere™ ODS-L80 0.95 1.43 2.94 2.78 0.92 * For use in C18 Column Brand Name selection when referencing USP Monographs which specify the characteristics/performance of the C18 Column used in the method. Choose a brand as close as possible to the referenced column values for the best chance of duplicating the analytical results of the method. 2 © 2003 Waters Corporation.Waters is a trademark of Waters Corporation. YMC ™ is a trademark of YMC ™ , Co.,Ltd Guide to YMC™ Part Numbering System Guide to YMC™ Part Numbering System The YMC™ part numbering system is designed to help identify the product length, and column inner diameter. These are followed by fitting codes for being ordered. This system: internal use by Waters. The system is set out schematically below: •embeds in the part number a description of the product that can be read by the user; Gel Code Hardware Code • standardizes the YMC™ part numbers worldwide; and, Chemistry Pore Size in Particle Particle size Length Internal Code nm Shape in µm in mm diameter in mm • simplifies computerized stocking and ordering. The table below will enable you to “read” your part numbers, and confirm Example: a semipreparative column with the part number that a part number you are about to use for ordering is exactly what you “OC12S052520WT” is in your stock. “Read” the part number as need. This should reduce ordering errors that can delay delivery of the cor- OC/12/S/05/25/20, and you can determine that this column is: rect column. Octyl 12nm (120Å) Spherical 5 µm 25 cm 20 mm I.D. (C8) pores media particles long Gel Code & Hardware Code The part numbers are composed of a gel code, which describes the pack- Please do not use this table to write your own part numbers. Certain attrib- ing in the HPLC column, and the hardware code, which describes the col- utes that are available in some stationary phases are not available umn itself. The gel and hardware codes are further divided into short codes in others. If you are unsure of a part number, please check the catalog or that describe the chemistry, pore size, particle shape, particle size, column price list, ask your technical representative directly to confirm availability. Chemistry Code Pore Size in Angstroms Particle Shape Particle Size in µm Length in mm Internal Diameter in mm ODS-A™:AA60Å: 06 Spherical: S 3 µm: 03 10 mm: 01 0.2 mm: G0 ODS-AM™:AM80Å: 08 Irregular: I 4 µm: 04 20 mm: 02 0.3 mm: H0 ODS-AQ™:AQ120Å: 12 5 µm: 05 23 mm: G3 0.5 mm: J0 ODS-AL™:AL200Å: 20 6 µm: 06 30 mm: 03 1.0 mm: 01 PolymerC18™:PC300Å: 30 5/15 µm (10P): 11 33 mm H3 2.0 mm: 02 J’sphere™ ODS-H80: JH 1000Å A0 15 µm: 15 50 mm: 05 3.0 mm: 03 J’sphere™ ODS-M80: JM Proprietary or 10/20 µm: 16 75 mm: L5 4.0 mm: 04 J’sphere™ ODS-L80: JL unspecified: 99 15/30 µm: 21 100 mm: 10 4.6 mm: 46 YMCbasic™:BA 30/50 µm: 41 150 mm: 15 6.0 mm: 06 Octyl (C8): OC 50 µm: 50 250 mm: 25 8.0 mm: 08 Phenyl: PH 75 µm: 75 300 mm: 30 10 mm: 10 Butyl (C4): BU 150 µm: A5 500 mm: 50 20 mm: 20 TMS (C1): TM 350 µm: C5 1000 mm: A0 30 mm: 30 Cyano: CN Undesignated: 99 1500 mm: A5 50 mm (1000psi): 51 Amino: NH 2000 mm: B0 50 mm (2000psi): 52 PA-gel™:PA Irregular: 70 mm (1000psi): 71 Polyamine II: PB 5/20 µm: 12 70 mm (2000psi): 72 Silica: SL 10/25 µm: 17 100 mm (1000psi): A1 PVA-Sil™:PV 10/40 µm: 25 100 mm (2000psi): A2 Diol (NP): DN 25/40 µm: 32 150 mm (1000psi): B1 Diol (SEC): DL 40/63 µm: 52 150 mm (2000psi): B2 Protein-RP™:PR 63/210 µm: A4 200 mm (1000psi): C1 Pro C4 BS Pro C8 OS Pro C18™ AS 200 mm (2000psi): C2 Cartenoid CT Pro C18RS™ RS ™ Hydrosphere C18 HS © 2003 Waters Corporation.Waters is a trademark of Waters Corporation. YMC™ is a trademark of YMC™, Co.,Ltd. 3 YMC™ Column Selection Guide Choosing an HPLC Column YMC-Pack™ High Performance HPLC Columns High Performance Silica Supports YMC™ provides an extensive selection of packed columns to address your Spherical packing for long column life: YMC™ spherical packing pro- separation needs. Numerous specialty bonded phases are available to vides high performance with long column lifetimes. Spherical particles pack solve the most difficult separation problems. more uniformly than “spheroidal” or irregular supports. The greater mechan- ical stability of the column bed eliminates premature voiding. • Extensive selection of bonded phases available in five different porosities High surface area: The ultrapure YMC™ spherical supports have very high porosity, providing increased surface area for greater retention and • Column diameters from microbore to process scale increased resolution. In preparative