What is the mobile phase used in the DIY Chemical Solutions HPLC Qualification Kits and how much is used - FAQ
April 7, 2012
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Date: 7-APRIL-2012   Last Updated: 7-JULY-2025

The " Do It Yourself", Chemical Solutions HPLC Qualification Kits are designed for ease of use and compatibility with most standard HPLC systems. The mobile phase formulation is optimized to support both isocratic and gradient system performance checks using NIST-traceable standards.

Mobile Phase Composition

  • 14:86 Acetonitrile : 0.1% Acetic Acid in Water (v/v)
    • To prepare the aqueous component: Add 1 mL of glacial acetic acid per liter of water to make a 0.1% solution.
    • Mix 140 mL of acetonitrile with 860 mL of the 0.1% acetic acid solution to make 1 liter of mobile phase.
Tolerance:    12%-16% in acetonitrile absolute composition.
                                                0.08%-0.12% acetic acid absolute composition.
 

This composition provides:

  • Low UV background, ideal for UV detector qualification
  • Stable baseline for both isocratic and gradient testing
  • Compatibility with a wide range of analytes and column chemistries

Volume Requirements

  • For isocratic system testing: Prepare 1 liter of mobile phase.
  • For gradient system testing: Prepare 2 liters to ensure sufficient volume for method development, equilibration, and multiple injections.

Additional Tips

  • Always use HPLC-grade solvents and freshly prepare the mobile phase to avoid contamination or degradation.
  • Filter and degas the mobile phase before use to prevent baseline noise and pump cavitation.
  • Store unused mobile phase in amber bottles at room temperature and use within 24–48 hours for best results.

📦 Click HERE for PQ and HSQ Kit™ ordering information and images.
 

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