HPLC Method Transfer Calculator

HPLC Method Transfer Equation:

\[ \text{Adjusted Time} = \text{Original Time} \times \left( \frac{\text{New Length}}{\text{Original Length}} \right) \times \left( \frac{\text{Original Particle}}{\text{New Particle}} \right)^{0.5} \]

Original Time:

min

Original Length:

New Length:

Original Particle Size:

µm

New Particle Size:

µm

Adjusted Time:

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1. What is HPLC Method Transfer?

HPLC Method Transfer involves scaling chromatographic methods when changing column dimensions or particle sizes while maintaining separation quality. This calculator helps adjust retention times for different column configurations.

2. How Does the Calculator Work?

The calculator uses the HPLC method transfer equation:

Where:

\( \text{Original Time} \) — Original retention time (min)
\( \text{Original Length} \) — Original column length (mm)
\( \text{New Length} \) — New column length (mm)
\( \text{Original Particle} \) — Original particle size (µm)
\( \text{New Particle} \) — New particle size (µm)

Explanation: The equation accounts for changes in column length and particle size, with particle size having a square root relationship due to its effect on efficiency.

3. Importance of Method Transfer

Details: Proper method transfer ensures method robustness, maintains separation quality, and allows optimization of analysis time and solvent consumption when switching between different HPLC systems or columns.

4. Using the Calculator

Tips: Enter original retention time in minutes, column lengths in millimeters, and particle sizes in micrometers. All values must be positive numbers greater than zero.

5. Frequently Asked Questions (FAQ)

Q1: Why is particle size raised to the power of 0.5?
A: Particle size affects column efficiency with a square root relationship in van Deemter equation, hence the 0.5 exponent for scaling.

Q2: Can this calculator be used for UHPLC transfers?
A: Yes, the same principles apply when transferring methods between HPLC and UHPLC systems with appropriate column dimension changes.

Q3: What about flow rate adjustments?
A: Flow rate should be adjusted proportionally to maintain linear velocity: New Flow = Original Flow × (New Length/Original Length) × (Original Particle/New Particle)²

Q4: Are there limitations to this equation?
A: This assumes similar column chemistry and temperature. Significant changes in stationary phase may require method redevelopment.

Q5: How accurate is this calculation?
A: Provides good estimates for isocratic methods. For gradient methods, additional considerations for gradient time scaling are needed.