Overview

Chemical Solutions™ HSQ™ and PQ™ Qualification Kits were developed to provide laboratories with a practical and consistent approach to HPLC performance qualification. To support reliable testing across a wide range of instrument platforms, the qualification procedures utilize a standardized mobile phase that is easy to prepare and compatible with typical HPLC hardware and UV detection systems.

The recommended mobile phase provides stable chromatographic performance, low background absorbance, and reproducible results for qualification activities.

Recommended Mobile Phase Composition

The standard qualification mobile phase consists of:  14:86 (v/v) Acetonitrile : 0.1% Acetic Acid in Water

Preparation of the Aqueous Component

Prepare a 0.1% acetic acid solution by adding:

• 1 mL glacial acetic acid
• To 1 liter of purified water

Mix thoroughly before use.

Preparation of the Final Mobile Phase

Combine:

• 140 mL Acetonitrile
• 860 mL of 0.1% Acetic Acid in Water

Mix thoroughly to obtain one liter of qualification mobile phase.

Acceptable Composition Tolerances

To accommodate minor preparation variability, the qualification method allows the following ranges:

Acetonitrile

Acceptable range:  12% to 16%

Acetic Acid

Acceptable range:  0.08% to 0.12%

Maintaining the mobile phase within these limits helps ensure consistent qualification performance.

Why This Mobile Phase Is Used

The selected formulation was designed to support a variety of qualification measurements while maintaining simplicity and reproducibility.

Benefits include:

• Low UV absorbance background
• Stable detector baselines
• Reproducible chromatographic performance
• Compatibility with qualification standards
• Suitability for both isocratic and gradient systems

These characteristics help support reliable qualification data and reduce the potential for mobile phase-related variability.

Mobile Phase Volume Requirements

The quantity of mobile phase required depends on the type of qualification testing being performed.

Isocratic Qualification Testing

Recommended preparation volume:  1 liter

This volume is generally sufficient for system equilibration and routine qualification testing.

Gradient Qualification Testing

Recommended preparation volume:  2 liters

The additional volume supports:

• Gradient equilibration
• Multiple injections
• System flushing
• Extended qualification procedures

Mobile Phase Preparation Best Practices

For optimal results:

• Use HPLC-grade solvents.
• Prepare mobile phases using calibrated volumetric equipment.
• Mix thoroughly before use.
• Filter when appropriate for laboratory procedures.
• Degas before placing the mobile phase on the instrument.

Proper preparation helps minimize:

• Baseline noise
• Pump cavitation
• Flow inconsistencies
• Detector instability

Storage Considerations

For maximum consistency during qualification testing:

• Prepare mobile phase as close as practical to the time of use.
• Store unused mobile phase in clean, suitable laboratory containers.
• Protect stored mobile phases from contamination.

Freshly prepared mobile phases are generally preferred for qualification activities.

Key Takeaways

• HSQ™ and PQ™ Qualification Kits use a standardized mobile phase consisting of 14% acetonitrile and 86% 0.1% acetic acid in water.
• The formulation supports both isocratic and gradient qualification testing.
• Low UV background and stable baselines help support detector qualification.
• One liter is typically sufficient for isocratic tests, while two liters are recommended for gradient evaluations.
• Proper mobile phase preparation and handling contribute to reliable qualification results.

For Chemical Solutions™ HSQ™ and PQ™ Qualification Kit Specifications, Qualification Resources, Product Images, and Ordering Information, view Chemical Solutions™ HSQ™ and PQ™ Qualification Kit Specifications, Qualification Resources, Product Images, and Ordering Information.