Date 15-OCT-2020 Last Updated: 6-June-2025
FAQ: Can I use the certified HPLC column supplied in the qualification kits for UHPLC performance qualification?
In most cases, no. The certified HPLC column included in the PQ™ and HSQ™ Qualification Kits is specifically designed and validated for standard HPLC systems, not for UHPLC platforms. However, thoeretically you do not need a sub 2um particle size stationary phase to qualify the instrument.
Why this matters:
- Column dimensions and particle size: The qualification column is typically packed with 5 µm particles, which are ideal for HPLC flow rates and pressures. UHPLC systems, by contrast, are optimized for sub-2 µm particles and operate at significantly higher pressures (up to 15,000 psi or more).
- System suitability: Using an HPLC column on a UHPLC system may not generate meaningful performance data. The system’s resolution, efficiency, and pressure response will not be properly challenged, potentially masking issues in pump performance, gradient formation, or detector response.
- Qualification integrity: For a valid UHPLC performance qualification, the test column must match the system’s design specifications. This ensures that metrics like plate count, peak symmetry, and retention time precision are evaluated under appropriate conditions.
Recommendation:
If you're qualifying a UHPLC system, we recommend using a dedicated UHPLC test column with appropriate particle size and pressure tolerance. Contact us for guidance on compatible columns and test protocols tailored for UHPLC platforms.
🔗 Click HERE for HSQ Kit ordering information and pictures.
The MICROSOLV HSQ Kit™ is supplied with a 5um particle size HPLC column that can be used to qualify a UHPLC instrument; you maynot need a 2um particle size in a UHPLC column.
PLEASE NOTE: Calculation of system parameters such as extra column dispersion may involve measurements of column related values such as efficiency but these values are relative to each other. For example, the earlier a peak elutes, the greater the extra column dispersion is relative to the peak volume itself. In an ideal system, all peaks would have about the same efficiency, so as soon as you see a trend of early peaks having lower efficiency, leading up to a constant efficiency with later eluting peaks, it is due to extra column dispersion. So it doesn't matter whether a 4um or 2um particle size column is used to determine the extra column dispersion value.