YMC USER'S MANUAL

Column Care and Use Instructions

Pro series
YMC-Pack Pro C18, Pro C18 RS, Pro C8, Pro C4 /Hydrosphere C18
1. Introduction
Thank you for purchasing a YMC high-performance liquid chromatography (HPLC) column “Pro series”.
Pro series, which are based on ultra high purity silica gel and adopted the advanced endcapping technology, provide
excellent separations with symmetrical peaks for a wide range of compounds, especially basic pharmaceuticals.
Pro series, which are manufactured under highly controlled conditions, must pass a series of stringent tests before being accepted
for shipment. (Please refer to the column inspection report). To ensure optimal performance and durability of the column, please readthese instructions carefully before using this column.

2. Specifications
Packing material
Pro C18
Hydrosphere C18
Pro seriesParticlesize (μm)

Pore size(nm)C%

3, 5, 101216

3, 51212

Pro C18 RS

3, 5822

Pro C83, 51210

Pro C4

3, 5127

Usable pH
range

Usable temperature range
Regular use
Upper(recommended)
limit

2–8
1 – 1020 – 40 °C50 °C

2 – 7.5

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 a mobile phase
containing buffer salts/additives, extra care must be taken to prevent salt precipitation.

4. Mobile phase
・ The correct direction of the solvent flow is indicated by an arrow on the column identification label.
・ Aqueous or non-aqueous solvent can be used as a mobile phase. Repetitive replacement among solvents with large differencein polarities might degrade the column performance. In general, organic solvents are acetonitrile, methanol and tetrahydrofuran(THF) are recommended for regular use. When using THF as a mobile phase, be mindful of the solvent resistance of yoursystem or tubing (PEEK parts are especially unsuitable for use with THF).
・ Recommendations of pH for column use are shown in the specifications table in section 2. When using the column at pH near
the upper or lower limit, the column lifetime will shorten under certain conditions by temperature and mobile phase composition.
In addition, please note the following.
For all products:

When using the column at pH near the limit, mobile phase containing 10% concentration of organic
solvent should be used.

For Hydrosphere C18: Although Hydrosphere C18 is compatible with 100% aqueous mobile phase, 10% concentration oforganic solvent should be contained when using the column at pH near the limit.
For Pro C18 RS:

When using the column at pH more than 9, mobile phase containing 50% concentration of organic
solvent should be used.

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(UM231220AE) 1/2

YMC USER'S MANUAL
・ When using Pro C18 RS column, its extremely high hydrophobicity might cause difficulties of replacement or equilibration withmobile phase containing low concentration of organic solvent. The mobile phase should be contained more than 10% of
methanol or 5% of other lower polar organic solvent. When replacing methanol aqueous solution with acetonitrile aqueoussolution, 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.

5. Column cleaning (general method)・ 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 column after using mobile phases not containing buffer salts/additives. Usable concentration oforganic 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.
・ When using mobile phase containing buffer salts/additives, first replace with a water/organic solution containing no buffer
salts/additives (A ratio of water to organic solvent should be set at the same proportions as a mobile phase). Then flush thecolumn in accordance with the method described above. Mobile phase containing about 50 mM or less in buffer salts/additives
can be replaced directly with 60% acetonitrile aqueous solution.
・ Flushing with 100% water after using the column around the pH limit might shorten the column lifetime. Flush the column withwater/organic solution as described above, such as 60% acetonitrile aqueous solution.
・ Once macromolecules such as proteins or polysaccharides are adsorbed onto the gel, they are hardly removed, even if solventswith 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.

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. For 3 μm particle size and 150 mm or 250
mm length column, the operating pressure should be kept under 30 MPa (4350 psi).
・ 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.

・ Recommendations of temperature for column use are shown in the specifications table in section 2. When using the preparative
columns at a higher temperature than ambient temperature, the thermal mismatch between mobile phase and column
temperature might cause peak distortion. To avoid those phenomena, we recommend preheating the mobile phase.

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(UM231220AE) 2/2