Waters HPLC To UPLC Gradient Calculator

Waters HPLC To UPLC Gradient Equation:

\[ Adjusted\ Gradient\ Time = Original \times \left( \frac{UPLC\ Particle}{HPLC\ Particle} \right)^{0.5} \]

Adjusted Gradient Time:

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1. What Is Waters HPLC To UPLC Gradient Calculator?

The Waters HPLC To UPLC Gradient Calculator helps chromatographers scale HPLC methods to UPLC methods by adjusting gradient times based on particle size differences. This ensures method transfer while maintaining separation efficiency and resolution.

2. How Does The Calculator Work?

The calculator uses the gradient scaling equation:

\[ Adjusted\ Gradient\ Time = Original \times \left( \frac{UPLC\ Particle}{HPLC\ Particle} \right)^{0.5} \]

Where:

\( Original \) — Original gradient time in minutes
\( UPLC\ Particle \) — UPLC column particle size in micrometers (µm)
\( HPLC\ Particle \) — HPLC column particle size in micrometers (µm)

Explanation: The equation accounts for the relationship between particle size and optimal flow rate/gradient time, ensuring equivalent linear velocity and separation quality when transferring methods between HPLC and UPLC systems.

3. Importance Of Method Scaling

Details: Proper method scaling is crucial for maintaining chromatographic performance when transferring methods between different instrument platforms. It ensures consistent retention times, resolution, and peak capacity while taking advantage of UPLC's higher efficiency and speed.

4. Using The Calculator

Tips: Enter original gradient time in minutes, UPLC particle size in µm, and HPLC particle size in µm. All values must be positive numbers. Typical HPLC particle sizes range from 3-5µm, while UPLC particles are typically 1.7-2.5µm.

5. Frequently Asked Questions (FAQ)

Q1: Why scale gradient times when switching to UPLC?
A: Smaller particle sizes in UPLC columns require shorter gradient times to maintain equivalent separation efficiency and prevent excessive backpressure.

Q2: What are typical particle sizes for HPLC and UPLC?
A: HPLC typically uses 3-5µm particles, while UPLC uses 1.7-2.5µm particles for improved efficiency and faster separations.

Q3: Does this scaling affect method validation?
A: Method scaling requires re-validation to ensure the transferred method meets all performance criteria and regulatory requirements.

Q4: Are there other factors to consider in method transfer?
A: Yes, including system dwell volume, column dimensions, flow rate adjustments, and detector settings must also be optimized.

Q5: Can this calculator be used for other scaling scenarios?
A: The same principle applies when scaling between different particle sizes within the same platform, but additional validation is recommended.