Physical

HPLC method transfer

Scale flow, injection, and gradient between columns, with void volume and dead time.

F₂ = F₁(d₂/d₁)²(dp₁/dp₂)
HPLC transferF₂ = F₁(d₂/d₁)²(dp₁/dp₂); Vi₂ = Vi₁(d₂²L₂)/(d₁²L₁)
Calculated live

New method: 0.579 mL/min, 1.389 µL inj, 3.6 min gradient

1

Scaled to keep separation: flow by bore² and particle ratio, injection and gradient by column volume.

Geometric transfer preserves separation only if stationary-phase chemistry, temperature, and mobile phase are unchanged. Re-verify resolution after transfer.

New flow
0.579 mL/min
New injection
1.389 µL
New gradient
3.6 min
New t0
0.203 min
HPLC method transfer, Aliquot