Column Care Instructions

PRIMESEP COLUMN OPERATION AND STORAGE.

Good column care requires little additional time and will extend the lifetime of your PRIMESEP HPLC columns. Column care for PRIMESEP brand of columns involves operating in a suitable mobile phase and storing PRIMESEP column in an appropriate solvent system.

Mobile Phases for Column Operation 

General Guidelines

  1. TRY TO AVOID USING ALCOHOLS WITH Primesep A, Primesep 100, Primesep 200, Primesep C and Primesep P columns as you can esterify carboxylic acid attached to the surface of silica. The esterification might significantly Affect reproducibility and retention time of some basic analytes.
  2. Do not use or store SIELC HPLC columns in pH outside of recommended ranges. SIELC Columns require buffered or acidified mobile phases to be present in the column at all times.\
  3. Do not store PRIMESEP HPLC columns in mobile phases containing modifiers that degrade easily such as triethylamine (TEA), tetrahydrofuran (THF), and trifluoroacEtic acid (TFA). TEA and TFA are more likely to become contaminated when stored at room temperature, and should be refrigerated during long storage. Contamination may change the chromatography or change the column. Use HPLC-grade solvents and additives for your chromatography.
  4. To achieve reproducible results, please use a thermostat. Operating temperature for all SIELC columns is 0-50°C
  5. To achieve reproducible results, always equilibrate SIELC columns prior to injection. In some cases replacement of the mobile phase inside the column does not make the column sufficiently equilibrated. SIELC columns’ stationary phase retain components of the mobile phase as counter ions. To equilibrate the column in this case requires a replacement of all the ions adsorbed in the column stationary phase. Typically, 50 mM buffer would equilibrate a column in 1 hour. If only the concentration of the buffer changed, the 2-4 volumes of the column are sufficient to reach and equilibration state. 
  6. It is important to operate column below the specified maximum pressure limits (5000 psi). If you exceed maximum pressure, the column can be destroyed. Leaking column is an indication of excessive pressure range.
  7. Limit the use of tetrahydrofuran to 10% with SIELC columns
  8. An arrow on the column label is used to specify direction of flow through the column. Typically you should operate the column in the direction marked. If column needs to be washed in the opposite direction use reduced flow rate in order not to create pressure which can disturb packing. 
  9. Use only fingers to tighten PEEK fittings to the column connection. 

  

Table 1. Operational pH Ranges for SIELC Columns

Column

pH Limits/

Recommended 

Operating Range

Short Term Storage Long Term Storage

Primesep A, Primesep 100, 

Primesep 200, Primesep C,

Primesep P, Primesep AB, 

Primesep SP, Promix AP,

Obelisc R

1.5-7.0/ 2.0-6.5

Buffered or acidified

mobile phases within

recommended pH range

ACN/water/formic or acetic acid = 75/25/0.1 

ACN/water/acetic acid

Primesep B, Primesep D

Primesep B2, Primesep B4,

Primesep SB, Promix MP,

Promix LP, Obelisc N

1.5-5.0/ 2.0-4.5

Buffered or acidified

mobile phases within

recommended pH range

ACN/water/formic or acetic acid = 75/25/0.1 

 

Other Recommendations:

Preform the column performance test using conditions described in  the Certificate of Analysis and compare with the result in the report. Due to variations in particular LC systems you may observe slightly different results.