YMC HPLC COLUMN USER'S MANUAL

Column Care and Use Instructions

YMC-GPC
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
Connect the GPC column in the mobile phase flow direction indicated and tighten
the 1/16 inch nut and ferrule using wrenches on the 1/16 inch nut and the actual end
fitting.
It is recommended that several drops of mobile phase have been pumped before
the column outlet is connected to another column or detector to clean out the end
fitting of any particulate matter which may be present.
To avoid loosening the end fittings and causing leaks, wrenches must be used on
the end fitting adjacent to the connecting nut and NOT on the column barrel or theopposite end fitting, See Figure 1.

Figure.1. Don’t use wrenches on the flats.

3. Recommended flow rate
The recommended flow rates are given in Table1, however, higher viscosity mobile phases should be used at reduced flow rates
or elevated temperature. Flow rates should be changed progressively and pressure pulses limited. At no times should the maximum
operating pressure of the column be exceeded (see Table 1).
Table 1. Recommended flow rate
Particle size
Column type(μm)

Flow rate(mL/min)

Recommended
flow rate (mL/min)

Recommended
pressure (MPa)

Pressure limit (MPa)YMC-GPC 21.2 mm I.D.

10

3.0 – 15.0

10.0

3.514YMC-GPC 40 mm I.D.

1030.0 – 40.0

35.0

3.0

144. Sample preparation and injection
If maximum resolution and expected column lifetime are to be achieved care must be taken in sample preparation.
To avoid blockage of the column frits, sample filtration is recommended (0.5 – 2.0 µm depending on molecular weight [MW]). A
guard column will further protect the columns with little detrimental effect on performance.
Optimum sample volumes and concentrations are best determined for each type of analysis and are dependent on sample MW.
Broad distribution polymers can generally be injected at higher concentrations than lower polydispersity samples. Overloading will notdamage the column, but distorted peaks and spurious results will be obtained.
Excessive injector loop volume will contribute to band broadening and reduce system performance. Injection volume
recommendations are shown in Table 2.
Table 2. Concentrations and injection volumes
Column type

Recommended concentration (%)

Recommended injection
volume (μL)

YMC-GPC 21.2 mm I.D.

0.5 – 5.0500 – 2000

YMC-GPC 40 mm I.D.

0.5 – 5.01000 – 5000

5. Mobile phase
YMC-GPC columns are compatible with an extensive range of organic solvents. Mixed organic solvent systems can also be used,
but water should not be used except at concentrations less than 10% by volume in a miscible organic mobile phase. All mobilephases should be of high purity and should be filtered and degassed prior to use. YMC-GPC columns are normally supplied in
ethylbenzene unless otherwise stated, and can be flushed directly from ethylbenzene to THF at 3.5 mL/ min. Unstabilized THF (forexample, HPLC grade) is not recommended as an mobile phase due to the attack of peroxide on the gels.

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YMC HPLC COLUMN USER'S MANUALYMC-GPC columns can be transferred to other mobile phases with no deterioration in performance. When transferring to anothermobile phase, miscibility and viscosity of the new mobile phase are of primary consideration. The transferring procedure is describedin Figure 2. When transferring to another mobile phase without heating, use a lower flow rate though it depends on viscosity of themobile phase. Note that loss of resolution or peak broadening might be seen if heating cannot be used when using high viscosity
solvents.
Column supplied : Ethylbenzene

Transfer to :HexanePerfluoroalkanes
Dichloromethane
MEK
Ethyl acetate
Chloroform

Transfer to :Dichloroethane
Cyclohexane

Transfer to :DMAC
Pyridine
DMF
DMSO
NMP

1. Flush column with acetone at （3.5 mL/min） or ＜20 mL/min＞ for 2 C.V.
2. Flush with new mobile
phase at （3.5 mL/min）or ＜20 mL/min＞ for 2C.V.

2. Flush with new mobile phase at （1.4 mL/min）or ＜7 mL/min＞ for 2 C.V.

3. Increase column
temperature to 60 °C
3. Operate column in new
mobile phase at required
flow rate
C.V. : Column Volume

3. Increase column
temperature to 70 °C

3. Operate column in new
3. Operate column in new
mobile phase at required flow
mobile phase at requiredrate
flow rate*Flow rate : （ ） 21.2 mm I.D. ＜ ＞ 40 mm I.D.
Figure 2. Mobile phase transfer guideTransfer to :o-DCB
TCBo-CPm-Cresol

1. Increase column
temperature to 50 °C
2. Flush with new mobile
phase at （0.7 mL/min）or ＜3.5 mL/min＞ for 2C.V.
3. Increase column
temperature to 100 –150 °C
3. Operate column in new
mobile phase at required
flow rate

6. Column testing
Every YMC-GPC column is supplied with a test certificate indicating the test conditions and the column performance.
Measurements of column performance are described below:
Efficiency (1/2 ht) (Plates/m) N = 5.54 (t/W1/2)2/L
Efficiency (5σ) (Plates/m) N = 25 (t/W5σ)2/L
Symmetry = a/bt:
W1/2 :
W5σ :
L:
a、b :

Peak elution time
Peak width at half peak height
Peak with at 4.4 % of peak height
Column length in meters
Peak widths either side of the perpendicular measured at 10 %of peak height.

Column efficiency is dependent on many experimental factors (system dead volume, mobile phase, flow rate, test probe,
temperature, and so forth) and test results may differ slightly from those quoted on the column certificate due to variability in theseparameters.

7. Storage
On removing the column from the system, the end plugs must be replaced to prevent the column from drying out by evaporation,
since subsequent shrinkage of the gel and disruption of the packing will occur. The end plugs need only be applied finger tight. All
mobile phases mentioned previously are suitable for storage, but unstabilized THF should not be used.

8. Maintenance
Deterioration in column performance may occur as a result of damage to the packed bed or as a result of blockage in the column
frits. In the case of frit blockage, the column can be reverse flushed at 1.0 mL/ min for 1 minute to remove loosely retained material.

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
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