• When the product is delivered to you, please check that there are no defects and damage.
• Please check that you have received the HPLC/UHPLC column that you have ordered. Check the product name, column size, connection type of Develosil series you have ordered.
• This packaging includes the following items.
  ・ Column ・ Test report ・ Certification of analysis
• Develosil series are managed with product numbers and batch numbers. Therefore, please keep the test report.

• Any shocks such as dropping and striking to the product may result in damage to its structure and performance.
• As a guideline column pressure use less than 20 MPa (2900Psi, 200bar).
• For Develosil HSR series = 25MPa (3625psi, 250bar).
• For Develosil HB series = 50MPa (7251psi, 500bar)

・ Before installing the column, replace the equipment and the piping with mobile phase.
・ Flow through according to the arrow marked on the column body.
・ Selection of pipe inner diameter greatly affects column performance, so please use appropriate tubing.
・When removing the column, please do after the pressure display becomes “0”.

Recommended column inner diameter and tube inner diameter

• Column is shipped in the test report. When feeding a mobile phase, make sure that the compatibility of the solvents as well as salt precipitation.
• Make sure to filtrate the mobile phase using a 0.45um membrane filter before using the column.
• When high water content mobile phase is used, extreme change in the retention time may be observed when the flow of the mobile phase is stopped. In such a case, wash the column with mobile phases containing 60% Acetonitrile or Methanol, and then repeat the analysis.
• As a guideline column temperature use less than 50℃.
• Sample should be dissolved in the mobile phase or solvent weaker than mobile phase, which helps avoid sample precipitation at the column inlet/head and inconsistent retention values.
• The Develosil series has a pH range of use, but the life of the column will change depending on conditions such as the type of buffer solution used, temperature, mobile phase composition, etc. For long-term, stable analysis it is recommended to analyze with a composition containing low concentration buffer solution, additives, and organic solvents. Please refer to “pH range used” for the pH range used.

Recommended type of buffer solution

Columns for Reverse-Phase
A. Column washing procedure：
-When using solvents containing buffer solutions or ion pair reagents, it is necessary to wash them so as not to leave them in the column. After use, wash with solvent of the same composition.
-Example: Analytical eluent- MeOH/25mM (NH4)2HPO4 (pH7.0) =50/50 Washing eluent- Methanol/Water=50/50Flush the column ten times the column volume at a reduced flow rate (approximately 50% lower than the normal flow rate).

B. Storing of columns：
-After washing, seal the column with the plugs provided and store at temperature stable place.

-For water 100% mobile phase conditions, replace with acetonitrile/waterer or methanol / water before storing. Please keep away from long-term storage only with water.

C. Applicable Columns:
ODS-K, ODS-N, ODS-P, ODS-T, ODS-A, TMS, C8, Ph, PhA, CN, NH2, ODS-HG, ODS-UG, ODS-MG, ODS-SR, PAHS, C30-UG, RPAQUEOUS, RPFULLERENE, Combi-RP, RPAQUEOUS-AR, TMS-UG, C8-UG, Ph-UG, CN-UG, XG-C18M, XG-C18LC, XG-C30M, XG-C1, XG-CN, HSR C18, HSR AQ C18, HSR C1, HSR C18 Peptide, 300 ODS-HG, 300 ODS-UG, 300 C8-HG, 300 C4-HG, ANIDIUS, HB series

Columns for Normal-Phase
A. Column washing procedure：
-When using additives such as acetic acid, it is necessary to wash them so as not to leave them in the columns. After use, wash with solvents of the same composition.
-Example: Analytical eluent- Hexane/Ethanol/Acetic acid＝99/1/0.1 Washing eluent- Hexane/Ethanol=99/1
-Flush the column ten times the column volume at a reduced flow rate (approximately 50% lower than the normal flow rate).
-It is effective to increase the ethanol content when the sample adsorption is strong.
B. Storing of columns：
-After washing, seal the column with the plugs provided and store at temperature stable place.
C. Applicable Columns
Columns specific to normal phase：30, 60, 100,
Columns that can be used for Normal-Phase: NH2, CN, CN-UG, XG-CN, 100 Diol, 300 Diol, ANIDIUS

Columns for Gel Filtration
A. Column washing procedure：
-High concentration of buffer solution may be used for gel filtration chromatography. After using, wash the column enough because acid and salt tend to remain.
-Example： Analytical eluent- 0.1M Buffer＋0.2M NaCl (pH6.8) Washing eluent- Pure water
-Flush the column ten times the column volume at a reduced flow rate (approximately 50% lower than the normal flow rate).

ii. Storing of columns：
-After washing, seal the column with the plugs provided and store at temperature stable place.
-When storing the column for a long period, replace with 0.05% sodium azide aqueous solution.

iii. Applicable columns
-100 Diol, 300 Diol

They can be also used with Normal-Phase (Develosil NH2, CN, CN-UG, XG-CN, 100 Diol, 300 Diol, ANIDIUS). When switching the eluent of columns, follow the procedure below.
Example Column: Develosil NH2
Replacing eluent from Acetonitrile/Water＝60/40 to Hexane/Ethanol＝ 99/1
１）Replace the eluent to Ethanol.
Flush the column ten times the column volume at a reduced flow rate to avoid sudden pressure increases.
２）Flush the column with Hexane/Ethanol＝99/1

Use the mutually miscible eluent such as ethanol when switching from Reversed- Phase to Normal-Phase.