YMC-Triart C18 ExRS 1.9 µm
Contents
1. YMC USER S MANUAL Column Care and Use Instructions YMC Triart C18 ExRS 1 9 um UHPLC Column 1 Introduction Thank you for purchasing a YMC Triart C18 ExRS 1 9 um column for ultra high performance liquid chromatography UHPLC YMC Triart C18 ExRS 1 9 um is a UHPLC column based on 1 9 um hybrid silica gel YMC Triart C18 ExRS 1 9 um exhibits higher efficiency and maintains efficiency over a wide range of flow rate and pressure compared to conventional 5 um 3 um columns Therefore YMC Triart C18 ExRS 1 9 um enables ultra high throughput analysis and remarkable decrease in analysis time Additionally YMC Triart C18 ExRS 1 9 um column offers easy method transfer between UHPLC and HPLC since it shows identical selectivity to conventional 5 um and 3 um YMC Triart C18 ExRS columns with the same bonded phase YMC Triart C18 ExRS 1 9 um column is manufactured under highly controlled conditions and must pass a series of strict tests before being accepted for shipment Please refer to the column inspection report In order to ensure optimal performance and durability of the column please read these instructions carefully before using this column 2 Specifications Particle Nene Usable temperature range Packing material size i Functional group Regular use H range um i recommended Bpeti j ee 70 C 3 Recommendations for column connections detector settings and data processing considerations The PT at the end of the product code in2. dicates the style of column endfittings See below for details Consideration of connector and endfittings Tubing and connector Column The end of the Port depth Style of endfittings ww code gt lt ca 2 mm 0 09 inch Parker style UPLC compatible Port depth UPLC is a registered trademark of Waters Corporation Tubing must have flat ends and must bottom out in the column endfitting Tubing must be connected to the column correctly to avoid creating a void between the column frit and tubing which can cause a leak and result in poor column performance e g peak tailing loss of theoretical plate number The 1 9 um column for UHPLC gives high operating pressure compared to conventional 5 um 3 um columns Particular attention should be devoted to the usable maximum pressure of tubing and LC system Generally a UHPLC system that has 60 MPa 8 700 psi or higher pressure tolerance is recommended for use with a 1 9 um column We have a connector with a re adjustable ferrule which provides flawless sealing at 137 MPa 20 000 psi Please inquire for related details The extra column volume has a great impact on band spreading To minimize the influence of band spreading on chromatographic performance especially when using columns in 2 0 mm I D and smaller the LC system should be optimized as described below gt The shortest possible length of tubing with narrow inner diameters tubing less than 0 15 mm 0 006 inch I D
3. is recommended should be used for the connection from the injector to the column and from the column to the detector Make sure not to have a void in the connection gt Use a detector equipped with low volume flow cell designed for the narrow bore column gt Usean injector for the narrow bore column and a low volume sample loop A sampling rate and a detector response time constant should be optimized to acquire more than 10 data points across a peak For UHPLC analysis using 1 9 um columns we recommend a sampling rate of about 10 points per second or higher and a detector response of 0 1 s or faster to detect the earliest eluting sharp peak propery 4 Shipping solvent 100 Acetonitrile Replace with this solvent for storage When replacing a mobile phase containing buffer salts additives extra care must be taken to prevent salt precipitation UM150724AE 1 2 YMC USER S MANUAL 5 Precautions for use The correct direction of the solvent flow is indicated by an arrow on the column identification label Do not disconnect a column from the LC system before the pressure drops to zero The column pressure limit and recommended flow rate are the following Particle size Pre einmit Column I D and recommended flow rate Mobile phase condition acetonitrile aqueous solution 2 0mm I D 0 2 0 8 mL min ee 3 0mm I D 0 4 1 6 mL min Avoid using a column repeatedly near the pressure limit or abrupt change in press
4. mn Usable concentration of organic solvent is up to 100 A cleaning solution containing THF might be effective when removing highly hydrophobic lipid soluble substances that are adsorbed onto the gel After using mobile phase containing buffer salts additives First replace with a water organic solution containing no buffer salts or additives A ratio of water to organic solvent should be set at the same proportions as a mobile phase Then flush the column in accordance with the method described above Mobile phase containing about 50 mM or less in buffer salts and additives can be replaced directly with about 60 acetonitrile aqueous solution General proposals Flushing with 100 water after using the column around the pH limit might shorten column lifetime Flush the column with water organic solution as described above such as 60 acetonitrile aqueous solution Once macromolecules such as proteins or polysacchandes are adsorbed onto the gel they are hardly removed even if solvents with high eluting capability are used To avoid contamination of the column by them conduct sample pretreatment carefully before introduction into the column Alternatively use a guard column YMC CO LTD YMC Karasuma Gojo Bldg 284 Daigo cho Karasuma Nishiiru Gojo dori Shimogyo ku Kyoto 600 8106 Japan TEL 81 75 342 4515 FAX 81 75 342 4550 URL www ymc co jp E Mail sales ymc co jp UM150724AE 2 2
5. ts will Make sure of the miscibility among the organic solvents and take care to prevent the precipitation of buffer salts additives to avoid overpressuring the column When possible the sample should be dissolved in a solvent that is of the same composition as the initial mobile phase Using a stronger solvent than the initial mobile phase for sample dissolution might result in distorted peak symmetry and degraded resolution To prevent exposure of the column to excessive pressure the mobile phase and sample solution should be filtered through a 0 2 um membrane or smaller to remove particulates When using YMC Triart C18 ExRS column its extremely high hydrophobicity might cause difficulties of replacement or equilibration with mobile phase containing low concentration of organic solvent The mobile phase should be contained more than 15 of methanol or 10 of other lower polar organic solvent When replacing methanol aqueous solution with acetonitrile aqueous solution mobile phase containing less than 20 of acetonitrile may result in irregularities in retention time or peak shapes In these cases first replace with 60 acetonitrile aqueous solution and then replace with the mobile phase 6 Column cleaning general method After using mobile phase not containing buffer salts additives Flush the column with solution containing a higher ratio of organic solvent for washing out the compounds that have a great capacity for retention in the colu
6. ure to prevent shortening of the column life Adjust the flow rate appropriately because the pressure changes depending on the column length temperature types of organic solvent etc Aqueous or non aqueous solvents can be used as a mobile phase Repetitive replacement among solvents with large difference in polarities might degrade the column performance In general acetonitrile methanol and tetrahydrofuran THF are recommended for regular use Please also see the recommendation above under alkaline mobile phase conditions When using THF as a mobile phase be mindful of the solvent resistance of your system or tubing PEEK parts are especially unsuitable for use with THF Recommendations of pH and temperature for column use are shown in the specifications table in section 2 In addition please note the following Column lifetime varies depending on conditions of use such as pH temperature and mobile phase composition In general usage at higher temperatures higher concentrations of buffer salts additives and lower concentrations of organic solvents can shorten the column lifetime When using the column under alkaline conditions for a long time a column should be used with a low concentration about 1 10 mM of organic buffer such as triethylamine or glycine at a lower temperature less than 30 C The mobile phase containing methanol is expected to give a longer column lifetime under alkaline conditions than other organic solven
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