What is the maximum injection volume I can use with the ACQUITY UPLC BEH C18 1.7µm 2.1x100mm column (186002352) without overloading it? - WKB106988

ENVIRONMENT

• Mass overload
• Column loading capacity
• ACQUITY UPLC BEH C18 1.7µm 2.1x100mm

ANSWER

For isocratic methods, a reasonable injection volume is between 1% and 3% of the empty column volume.

For gradient methods, increased injection volume is possible if analytes fully retain at the head of the column before the gradient begins.

ADDITIONAL INFORMATION

For this column, the empty column volume is 0.346 mL.
1% = 0.00346mL (3 µL)
3% = 0.1038mL (10 µL)

Mass overload must also be considered. 
The loading capacity of this column for small molecules might be about 1.0 mg per injection. (This is only an estimate; it must be determined experimentally.)
The loading capacity of this column for peptides will be significantly less per injection in most cases, determined experimentally.