YMC USER'S MANUAL

Column Care and Use Instructions

YMC-Pack Diol-NP
YMC-Pack PVA-Sil
1. Introduction
Thank you for purchasing a YMC high-performance liquid chromatography (HPLC) column. YMC HPLC columns, which are
manufactured under highly controlled conditions, must pass a series of stringent tests before being accepted for shipment. (Pleaserefer to the column inspection report). To ensure optimal performance and durability of the column, please read these instructions
carefully before using this column.

2. Column connections
The “WT” or “PTH” at the end of the product code indicates the style of column endfittings.
WT = Waters style / PTH= Parker style

3. Shipping solvent
Indicated in the COLUMN INSPECTION REPORT. Replace with this solvent for storage. When replacing mobile phases, makesure of the miscibility among the solvents. When the displacement from non-aqueous to aqueous solvent or vice versa, flush thecolumn with more than 10 column volume of compatible solvent like 2-propanol and THF.

4. Mobile phase
In general, the column is used as normal phase. The column can be used as Hydrophilic Interaction Chromatography (HILIC). We
recommend using the same column exclusively for separation mode for column lifetime.
【Normal phase】
・ In general, alkane (n-hexane or n-heptane), alcohols (methanol, ethanol, 2-propanol), ethyl acetate, dichloromethane, chloroform,
are recommended for regular use.
・ Addition of alcohol to alkane is basic. Acetonitrile, tetrahydrofuran (THF), dioxane etc. are also usable. When preparing mobile
phases, make sure of the miscibility among the solvents.
・ When a target compounds is ionic, addition of modifier at trifluoroacetic acid (TFA), acetic acid etc. can improve peak shape andseparation reproducibility. High concentrations of modifiers can result in reducing column lifetime. In this case, the same column
is not recommended for developing a new method, because exposure to acid or alkali may change the retention characteristics
of a column.
【HILIC】
・ The most suitable mobile phase is acetonitrile/water or buffer (approx. 90/10 – 60/40). In addition, general water-miscible organicsolvents.
・ Ammonium acetate or ammonium formate is most recommended as a buffer salt. Usually, 10 – 20 mM final salt concentration in amobile phase is sufficient. Depending on separation or solubility, adjustment in the range of 5 – 200 mM can be made.
【For all products】
・ The correct direction of the solvent flow is indicated by an arrow on the column identification label.
・ Non-aqueous or aqueous solvent can be used as a mobile phase. Repetitive replacement among solvents with large difference inpolarities might degrade the column performance.
・ When the displacement from non-aqueous to aqueous solvent or vice versa, flush the column with more than 10 column volume ofcompatible solvent like 2-propanol and THF.
・ Recommended pH ranges of the column are between 2 – 7.5. For PVA-Sil, pH ranges of 2 – 9.5 are available. Column lifetime isstrongly dependent on pH, temperature, and mobile phase composition. In general, high temperature and concentration of buffer oradditive, low organic solvent concentration will shorten column lifetime(UM230123GE) 1/2

YMC USER'S MANUAL

5. Column cleaning (general method)【Normal phase】
・ Flush the column with 2-propanol etc.
・ Replace with the mixture of n-hexane and alcohol etc. for storage.
【HILIC】
・ Use the mixture of organic solvent and water with a higher solvent strength than the mobile phase, such as acetonitrile/water (50/50),
to remove strongly retained substances. Usually, a solvent consisting of 50% water is sufficient to remove polar contaminants. Iffurther cleaning is required, flush the column with acetonitrile/water (5/95).
・ Once macromolecules such as proteins or polysaccharides are adsorbed onto the gel, they are hardly removed, even if solvents withhigh 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.
・ Replace with the mixture of acetonitrile etc. for storage.

6. Other environments
・ The operating pressure should be kept under 20 MPa (2900 psi) for 150 mm length column or less, under 25 MPa (3625 psi) for
250 mm length column, under 10 MPa (1450 psi) for 10 mm I.D. column or more.
・ To prevent exposure of the column to excessive pressure, the sample solution should be filtered through a 0.2 μm membrane or
smaller to remove particulates. We recommend using a pre-column filter to prevent the column frit from being clogged withsamples.
・ Avoid using a column repeatedly near the pressure limit or abrupt change in pressure 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.
・ The upper limit of column temperature is 50 °C. However, we recommend using the column at 20 – 40 °C, because column
lifetime varies depending on conditions such as pH.

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(UM230123GE) 2/2