YMC USER'S MANUAL

Column Care and Use Instructions

BioPro IEX Column, Accura BioPro IEX Column
for Separation of Proteins, Nucleotides, and Other Biomolecules
1. Introduction
Thank you for purchasing a YMC high-performance liquid chromatography (HPLC) column for ion exchange chromatography.
BioPro IEX Column and Accura BioPro IEX Column are based on a hydrophilic polymer bead with a strong-anion exchanger(quaternary ammonium group) or a strong-cation exchanger (sulfopropyl group). A porous-polymer type [BioPro IEX QA/SP] forhigh-performance and high-binding capacity and a non-porous-polymer type [BioPro IEX QF/SF] for high-throughput andhigh-resolution analysis are available.
BioPro IEX Columns and Accura BioPro IEX Columns, which are manufactured under highly controlled conditions, must pass aseries 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 read these instructions carefully before using this column.

2. Recommendations for column connections, detector settings, and data processing considerations
・ “WP” or “PTC” at the end of the product code indicates the style of column endfittings (see below for details).
Consideration of connector and endfittings
ColumnTubing and connector

Port depth
Item

The end of the product code

Column hardware specifications

Port depth

Style of endfittings

BioPro IEX

WP

PEEK

ca. 3 mm / 0.13 inch

Waters style (※)

Accura BioPro IEXPTC

Bioinert coated stainless steel

ca. 2 mm / 0.09 inch

Parker style※Use of a metal connector is not recommended. Inside parts of the column might be damaged if the metal connector is used.
・ 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 shortest possible length of tubing with narrow inner diameters (tubing less than 0.15 mm, 0.006 inch I.D. 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 agap in the connection.
・ A sampling rate and a detector response (time constant) should be optimized. When using BioPro IEX QF/SF and Accura BioPro
IEX QF/SF for ultra-fast separation, we recommend a sampling rate of about 10 points per second or higher and a detector
response of 0.5 s or faster to detect the sharp peak properly.
・ The correct direction of the solvent flow is indicated by an arrow on the column identification label.

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(UM240412AE) 1/3

YMC USER'S MANUAL

3. Recommendations for column use
・ Recommendations of conditions for column use are shown in the specifications table below. Avoid using a column repeatedly
near the pressure limit, or with abrupt change in pressure to prevent shortening of the column life.

Item

品種Particle size

QA / SP

5 µmColumn size
I.D.
Length(mm)
(mm)
4.6

Recommendedflow rate
(mL/min)

Flow rate limit(mL/min)Pressure limit
(MPa)

30

0.5–0.8

1.0

2.550

0.5–0.7

0.8

3.0

100

0.4–0.5

0.6

3.5

0.5–1.0

1.5

0.5–0.8

1.0

0.5–1.0

1.0

0.5–0.6

0.6

30
BioPro IEX

5 µm

4.650
100

QF / SF

30
3 µm

4.650
10050

4.65 µm

1001501.5
0.5–1.0

2.1
QF / SF100

0.1–0.250
10050
2.1

100150

12.0

25.0

10.0

1.0

18.0

0.3

10.0

0.2
1.5

0.5–1.0150

3 µm

4–60℃

30.0150

4.66.0
10.0

12.0

250
50

AccuraBioPro IEXUsabletemperature
range

1.0

0.1–0.2

0.2

0.1

0.1

12.0
18.0

4–80℃

25.0
25.0
30.0
15.0
15.0
20.0

4. Mobile phase and sample solvent
・ Shipping solvents are listed in the table below. They are the same ones as the mobile phases indicated in the “COLUMNINSPECTION REPORT”. When columns are not used for a long time, keep them in a cool place after replacing with the shipping
solvent.
Shipping solvent
BioPro IEX QA／QF
Accura BioPro IEX QF

20 mM Tris-HCl buffer (pH 8.1)

BioPro IEX SP／SF
Accura BioPro IEX SF

20 mM sodium phosphate buffer (pH 6.8)

・ Generally, samples are adsorbed onto the top of the column with 20 – 50 mM of buffer as the first mobile phase, then eluted witha salt-concentration gradient method (sodium chloride concentration commonly adjusted between 0 to 0.5 M) or pH-gradientmethod. We recommend flushing the column with buffer containing about 1 M of sodium chloride for each run to remove residual
impurities from the column with the final mobile phase.
・ Adjust the pH of the mobile phase in the range of 2 - 12.
・ Water-soluble organic solvent (maximum of 30%). Before adding such solvent, make sure that salt in the buffer will not precipitate.
Other additives such as urea (≤8 M) or guanidine hydrochloride (≤6 M), which are commonly used as protein denaturants,

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(UM240412AE) 2/3

YMC USER'S MANUAL
nonionic surface-active agents, cationic surface-active agents (limited to BioPro IEX QA/QF and Accura BioPro IEX QF), oranionic surface-active agents (limited to BioPro IEX SP/SF and Accura BioPro IEX SF) are usable.
・ Avoid solvents containing oxidant for the mobile phase.
・ When possible, the sample should be dissolved in a solvent that is the same composition as the initial mobile phase. Using adifferent buffer salts/additives concentration or a different pH solvent from the initial mobile phase for sample dissolution mightresult in decreased binding capacity and/or distorted peak shape.
・ To prevent exposure of the column to excessive pressures, the mobile phase and sample should be filtered through a 0.2 – 0.5
μm membrane filter. We recommend using a pre-column filter.

5. Column cleaning
A change of retention time or peak shape, and/or pressure increase might result from the adsorption of fat-soluble substances or
precipitated impurities in a sample. In such cases, follow these steps for column cleaning and regeneration. If these procedures will
not solve the problem, then we recommend that you use a new column.
【Cleaning procedures】
1. Replace mobile phase with the shipping solvent.
2. Then inject 4 – 5 mL of following cleaning solutions A) – D) while running the shipping solvent. The cleaning is recommended tobe conducted step-by-step and started from A). Column performance should be confirmed after cleaning with each solution. It
does not need to proceed to another cleaning solution if one solution can restore the column performance. We recommend usinga large-size sample-loop (≥2 mL).
Cleaning solutionA) 0.2 M NaOH aq／Acetonitrile（80／20）
B) 1 M Acetic acid aqC) Nonionic surfactant (like 0.02% BrijTM 35) in the shipping solventD) 6 M guanidine hydrochloride in the shipping solvent

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(UM240412AE) 3/3