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LC-TRANS-PRO v3.2
HPLC & UHPLC Advanced Method Transfer Suite
1. Column & Instrument Geometries
Scale dimensions geometricallyOriginal Column
Current RunTarget Column
Target RunLinear velocity compensation adjusts the target flow rate dynamically relative to the smaller particle size to maintain chromatographic efficiency.
2. Gradient Profile Translator
Scale time segments proportionally| Step | Original Time (min) | % B (Mobile Phase B) | Translated Time (min) | Action |
|---|
Interactive Profile Preview
3. Mobile Phase & Fluid Estimation
Transferred System Deliverables
USP <621> Compliance Status
1. Volumetric Injection Scale Factor
To conserve separation resolution, the analyte band width introduced into the column must scale directly with column volume:
2. Flow Rate & Velocity Conservation
Normalizing the chromatographic linear velocity prevents loss of peak efficiency. In sub-2 µm transfers, scaling can compensate for particle changes using:
Pure geometric scaling without particle compensation drops the term (d_p₁ / d_p₂).
3. Gradient Time Scaler (t_g)
To conserve elution selectivity in gradient separations, gradient step durations must be adapted to keep the gradient volume ratio matched relative to the column volumes:
4. Dwell Volume (V_D) Offset Calculations
System delay volumes introduce pre-gradient column washes. Transferring methods from high dwell volume instruments (HPLC) to ultra-low volume UHPLC requires a delay time injection offset:
A negative value indicates the gradient program must start prior to injection, or an isocratic hold is injected dynamically on the newer target platform.
5. USP Guidelines <621> Compliance
To prevent method revalidation, USP dictates that the ratio of column length to particle size (L/d_p) must remain within limits:
- Isocratic & Gradient: The L/d_p ratio must remain within a range of -25% to +50% of the original columns ratio.
- Alternatively, the equivalent theoretical plate count (N) change must fall within the same spectrum of original tolerances.