YMC USER'S MANUAL

Column Care and Use Instructions

YMC-Triart Diol-HILIC, Accura Triart Diol-HILIC
HPLC: 5 µm, 3 µm / UHPLC: 1.9 µm
1.

Introduction
Thank you for purchasing a YMC high-performance/ultra-high-performance liquid chromatography (HPLC/UHPLC) column. Triart
Diol-HILIC is a Hydrophilic Interaction Chromatography (HILIC) column based on 1.9 μm, 3 μm and 5 μm hybrid silica gel bondedwith dihydroxypropyl group. Triart Diol-HILIC is able to retain and separate highly polar compounds that are not retained in reversed
phase chromatography, with a mobile phase containing high concentration of organic solvent.
YMC-Triart Diol-HILIC and Accura Triart Diol-HILIC columns, which are manufactured under highly controlled conditions, must
pass a series of strict tests before being accepted for shipment (Please refer 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 hardware specifications

Column tubingColumn frit

YMC-TriartAccura Triart

YMC-Triart metal-free PEEK-lined

stainless steel

bioinert coated stainless steel

PEEK-lined stainless steel
PEEK

3. Recommendations for column connections
The "PT", “PTH”, "PTC", "PTP", or "WT" at the end of the product code indicates the style of column endfittings (see below fordetails).

Consideration of connector and endfittings
Tubing and connector

ColumnThe end of
the product codePort depth

Style of endfittings

PT/PTH/PTC/PTP

ca. 2 mm / 0.09 inch

Parker styleWT

ca. 3 mm / 0.13 inch

Waters style

Port depth

Wetted part (Inner surface of column tube) of “PTP” type column is PEEK. Its external tube is made of stainless steel. A special
care has to be taken to connect this “PTP” type column to your system. Refer to the “Instructions for column connection
YMC-Triart metal-free PEEK-lined”. https://www.ymc.co.jp/data/download/metal_free_column_connection_e.pdf

・ 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 µm column for UHPLC gives high operating pressure compared to conventional 5 µm/3 µm columns. Particularattention 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 µm column. We have a connector with are-adjustable ferrule, which provides flawless sealing at 137 MPa (20,000 psi). Please inquire for related details.

4. Shipping Solvent
Acetonitrile/water (90/10). Replace with this solvent for storage. When replacing a mobile phase containing buffer salts/additives,
extra care must be taken to prevent salt precipita.

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(UM230912BE) 1/3

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.
Column I.D. and recommended flow rate*2(Mobile phase condition : acetonitrile/aqueous solution)

Particle size

The end of theproduct code

Pressure limit*1

1.9 μmPT / PTP / PTC

100 MPa

2.0 mm I.D. :3.0 mm I.D. :0.2 – 0.8 mL/min
0.4 – 1.6 mL/min

PTH / PTP / PTC

45 MPa*3

2.1 mm I.D. :3.0 mm I.D. :4.6 mm I.D. :0.2 mL/min
0.4 mL/min
1.0 mL/min

5 μm, 3 μm

Column length of 150 mm and less: 20 MPa
2.0 mm I.D. :0.2 mL/min
Column length of 250 mm:
25 MPa3.0 mm I.D. :0.4 mL/min
Inner diameter of 10 mm and larger: 10 MPa
4.6 mm I.D. :1.0 mL/min*1 Avoid using a column repeatedly near the pressure limit or abrupt change in pressure to prevent shortening of the column life.
*2 We recommended using PTH / PTP / PTC type columns at a pressure of 30 MPa or less for regular use. Continuous use of the column near
the pressure limit may result in short column lifetime.
*3 Adjust the flow rate appropriately because the pressure changes depending on the column length, temperature, types of organic solvent etc.
WT・ Recommendations of pH and temperature for column use are the following.
Usable Temperature range
Usable pH range
Regular use (recommended)
Upper limit
pH 2 – 1020 – 40 °C

50 °C

 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 thecolumn 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 most suitable mobile phase is acetonitrile/water or buffer (approx. 90/10 – 60/40). In addition, general water-miscible organic
solvents such as described below can be used. Contrary to reversed phase separation, in HILIC separation, the lower polarityof mobile phase and the higher concentration of organic solvents will increase the retention. It is recommended to use a mobilephase containing at least 3% of aqueous solution to enhance the separation reproducibility by forming a stable hydrated layeron the surface of packing material.
Usable solvents and solvent strength (from weakest to strongest):
tetrahydrofuran（THF）＜acetonitrile＜2-propanol＜ethanol＜methanol＜water 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).

・ Ammonium acetate or ammonium formate is most recommended as a buffer salt. Usually, 10 – 20 mM final salt concentration
in a mobile phase is sufficient. Depending on separation or solubility, adjustment in the range of 5 – 200 mM can be made. Incase of gradient elution, each mobile phase composition should be adjusted to maintain a constant salt concentration during ananalysis. Before using or replacing a mobile phase, please confirm precipitation of salts is not observed. Avoid phosphate salt
and other low solubility buffers to organic solvents.
・ When possible, the sample should be dissolved in a solvent that is of the same composition as the initial mobile phase. Using astronger solvent than the initial mobile phase for sample dissolution might result in distorted peak symmetry and degraded
resolution. Sample should be injected after carefully checking the miscibility with the mobile phase to avoid precipitation ofsolutes or salts contained in the sample solvents.
・ To prevent exposure of the column to excessive pressure, the mobile phase and sample solution should be filtered through a
0.2 µm membrane or smaller to remove particulates.
・ 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.
 The shortest possible length of tubing with narrow inner diameters (tubing less than 0.15 mm, 0.006 inch I.D. isrecommended) 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.
 Use a detector equipped with low-volume flow cell designed for the narrow bore column.
 Use an injector for the narrow bore column and a low-volume sample loop.

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(UM230912BE) 2/3

YMC USER'S MANUAL
・ A sampling rate and a detector response (time constant) should be optimized to acquire more than 10 data points across apeak. For UHPLC analysis using 1.9 µm columns, we recommend a sampling rate of about 10 points per second or higher anda detector response of 0.1 s or faster to detect the earliest eluting sharp peak properly.

6.

Column cleaning (general method)・ 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 polarcontaminants. If further 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 ifsolvents 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 Karasuma-Gojo Bldg., 284 Daigo-cho, Karasuma Nishiiru Gojo-dori, Shimogyo-ku,
Kyoto 600-8106, JapanTEL +81-75-342-4515 FAX +81-75-342-4550
URL:www.ymc.co.jp E-Mail:sales@ymc.co.jp(UM230912BE) 3/3