Go Back Advanced Search Page

Column Cleaning Procedures

 

When a column is deteriorated by the foreign substances remaining in the column
or adsorbed by the packing material, the substances might be washed out by the
procedure described below. However, sometimes the procedure is not suffcient to
regenarate the column and, in such case, it is necessary to replace the column
with a new one. And, even if the column can be regenarated, the column performance
may be not so good as before. After cleaning the column, be sure to replace the
in-column solvent to the normal solvent. The use of a guard column at the up-stream
of the main column is recommended to prevent the main column from the deterioration
by the foreign substances. When a guard column is used, first, the guard column
will be deteriorated by the foreign substances and it is possible to prevent the
main column from the deterioration by replacing the guard column regularly.

When cleaning a guard column, be sure to disconnect the main column before
the cleaning.
If a guard column and a main column are
connected in series and the cleaning procedure
is performed, foreign substances in the guard
column may move to the main column and, as
the consequence, the main column may be deteriorated.

Category Column name Coulmn cleaning procedure
GPC
Clean-up
CLNpak EV Reverse the column on the HPLC system,
flow the eluent at 80% of ordinary flow rate. This is effective for the
foreign substances remaining in the column, however not effective for the
substances adsorbed to the packing material.
CLNpak PAE
Reversed Phase,

Hydrophilic Interaction

and Normal Phase

(Polymer-based)

Asahipak ODP, C8P, C4P Flow 50mL of dioxane, CH3CN
or methanol.
RSpak RP18, DS
RSpak DE, GOLF

CARB

RSpak DM-614 Flow 50mL of methanol, CH3CN
or H2O.
RSpak NN Inject 20 to 50 µL of 2N nitric acid aqueous
solution two or three times.
Asahipak NH2P-50 4E (Standard cleaning)
Reverse the direction and flow rate is 0.5mL/min.
1) H2O 10min, 2) 0.1M NaOH aq. 120min, 3) H2O 20min, 4) Eluent 60min
(Cleaning metal ions)
Reverse the direction and flow rate is 0.5mL/min.
1) H2O 10min, 2) 0.1M HClO4 aq. 60min, 3) H2O 10min, 4) 0.1M NaOH aq. 120min, 5) H2O 20min, 6) Eluent 60min
Reversed Phase,

Hydrophilic Interaction

and Normal Phase

(Silica-based)

ODSpak F Flow 50mL of dioxane, THF, CH3CN
or methanol.
Silica C18M, C18P
Silicapak E-411 Flow 50mL each of n-hexane, dichloroehtane,
methanol, dichloroethane and n-hexane.
SIlica 5SIL
Silica 5C8, 5C4,
5CN
Flow 50mL of dioxane, CH3CN
or methanol.
Silica 5NH There is no appropriate washing method.
Silica 5NPE
Silica 5PYE
Chiral
Separation
ORpak CDA, CDB, CDC There is no appropreate cleaning method.
ORpak CDBS, CRX There is no appropreate cleaning method.
Ion Exchange AXpak WA-624 Inject 1 to 2 mL of 0.1M NaOH aqueous
solution or 30% acetic acid aqueous solution. Connect the column in the
reverse way and flow the eluent at 0.5mL/min.
CXpak P-421S
IEC QA, DEAE

SP, CM

IEC DEAE3N-4T

SP-420N

Inject 500 µL of 0.1M NaOH aqueous solution,
50% formic acid aqueous solution, 20% CH3CN aqueous
solution or 20% dimethyl sulfoxide aqueous solution several times. Flow
the eluent at 0.3 to 0.5mL/min.
Asahipak ES-502N Flow aqueous solution of high salt concentartion
or 10mM NaOH aqueous solution for ionic adsorption and 50mM buffer solution
containing 50% CH3CN or methanol for hydrophobic
adsorption.
Asahipak ES-502C Flow aqueous solution of high salt concentartion
or 0.1N acetic acid aqueous solution for ionic adsorption and 50mM buffer
solution containing 50% CH3CN aqueous solution or methanol for
hydrophobic adsorption.
Hydrophobic HIC PH-814 Reverse the column on the HPLC system, flow the
eluent at 0.5mL/min and inject 1 to 2 mL of 0.1M NaOH or 30% acetic acid
aqueous solution several times.
Affinity AFpak 31 kinds Reverse the column on the HPLC system, flow an
appropriate eluent at 0.5mL/min.
Ligand Exchange SUGAR SC1011, 1821
SC1211
( for SC1011, 1821)

Flow 50mL of 0.1M Ca(NO3)2 aqueous solution at 50°C, 0.5mL/min.

(for SC1211)

After replacing in-column solvent completely with H2O,
flow 30mL of 0.1M Ca(NO3)2 aqueous solution at 50°C, 0.5mL/min. Then, replace the in-column solvent completely with
H2O and gradually increase the organic solvent
concentration to the concentration to be used.

SUGAR SP0810 Flow of H2O at 70 – 95 °C, 0.5mL/min. Then inject of 50µL of 0.2M Pb(NO3)2 aqueous 3 or 4 times.
SUGAR KS-800 Flow 10mM NaOH aqueous solution at 0.5mL/min.
After flowing the solvent, replace the in-column solvent completely with
H2O. (Sometimes, injection of 40µL of 0.1M
NaOH aqueous solution is will be sufficient.)
RSpak DC-613 Flow 50mL of Methanol, Ethanol, CH3CN, or 5mM NaOH aqueous solution. Or, inject 20 to 50 µL of 0.5M NaOH aqueous solution.
SUGAR SZ5532 After replacing in-column solvent completely
with H2O, flow 30mL of 0.1M Zn(NO3)2 aqueous solution at 50°C, 0.5mL/min. Then, replace the in-column solvent completely with
H2O and gradually increase the organic solvent
concentration to the concentration to be used.
USPpak MN-431 Flow 50mL of 0.1M Ca(NO3)2 aqueous solution at 50°C, 0.5mL/min.
Ion Exclusion SUGAR SH1011,1821 Flow 50mL of 25mM H2SO4 aqueous solution at 50°C, 0.5mL/min.
RSpak KC-811 Flow 50mL of 25mM H2SO4 aqueous solution at 50°C, 0.5mL/min.
Ion Chromatography IC I-524A –Pollution by accumulation of metals–

30minutes: 0.01M tartaric acid (pH3 to 4, adjust with 0.1M NaOH), 1.0mL/min

–Pollution by proteins–

150 minutes: 20% acetic acid aq., 0.2mL/min

150 minutes: water, 0.2mL/min

–Pollution by organics–

150 minutes: 20% acetonitrile/water, 0.2mL/min

150 minutes: water, 0.2mL/min

IC NI-424 Flow the mixture of eluent/methanol=90/10 at
0.6mL/min for two hours.
IC Y-521 Inject 100µL of 1M HNO3 aqueous solution several times.
IC YK-421 Flow 50mL of 50mM tartaric acid aqueous solution
or 50mM tartaric acid aqueous solution/CH3CN=50/50
at 0.3mL/min.
IC YS-50 Flow 50mL of 4mM Methanesulfonic acid aqueous solution
/CH3CN=70/30
at 1.0mL/min.
IC SI-90 –Pollution by low valency hydrophilic ions–

1. 15 minutes: deionized water

2. 60 minutes: 10 times concentrated eluent

3. 15 minutes: deionized water

4. 60 minutes: eluent

–Pollution by high valency hydrophobic ions–

1. 15 minutes : deionized water

2. 10 minutes: 5% acetonitrile aqueous solution

3. 60 minutes: 100% acetonitrile

4. 30 minutes: deionized water

5. 60 minutes: eluent

IC SI-50 –Pollution by low valency hydrophilic ions–

(flow rate 0.3mL/min)

1. 25 minutes : deionized water

2. 100 minutes : 10 times concentrated eluent

3. 25 minutes : deionized water

4. 100 minutes : eluent

–Pollution by high valency hydrophobic ions–

(flow rate 0.3mL/min)

1. 25 minutes : deionized water

2. 20 minutes : 5% acetonitrile aqueous solution

3. 100 minutes : 100% acetonitrile

4. 50 minutes : deionized water

5. 100 minutes : eluent

IC T-521 Flow 0.1M NaOH aqueous solution at
0.5mL/min for one hour and 0.1M Nitric acid aqueous solution at 0.5mL/min for one hour.
IC R-621
Aqueous SEC (GFC) OHpak SB-800 HQ
Reverse the column on the HPLC system,
flow the eluent at 50% of ordinary flow rate. This is effective for the
foreign substances remaining in the column, however not effective for the
substances adsorbed to the packing material.
SB400
PROTEIN KW-800
KW400
Maltimode Asahipak GS-HQ
Aqueous / Organic SEC Asahipak GF-HQ
Organic SEC
(GPC)
GPC KF-800
KF-400HQ
KF-600
K-800
KD-800
LF
HFIP-800
HFIP-600
HT-800
UT-800
AT-806S