Development of a Standard-Free Analytical Quantitative Method Based on Liquid Chromatography Using Relative Molar...

Area of Science:

• Analytical Chemistry
• Food Science
• Pharmacognosy

Background:

• Accurate quantification of chlorogenic acid (5CQA) in food products like apple juice is crucial.
• Traditional methods require analyte-specific analytical standards, which can have issues with purity, stability, and hygroscopic properties.
• The relative molar sensitivity (RMS) method offers an alternative by using a reference compound.

Purpose of the Study:

• To validate the robustness and reliability of the RMS method for quantifying 5-O-caffeoylquinic acid (5CQA) in apple juice.
• To demonstrate the utility of using caffeic acid (CA) as a stable reference compound in the RMS method.
• To compare the RMS method with traditional calibration curve methods for 5CQA quantification.

Main Methods:

• A collaborative study involving 10 laboratories was conducted.
• A standardized operating procedure for method validation was developed and implemented.
• The RMS value of 5CQA relative to caffeic acid (CA) was determined.
• 5CQA in five apple juice samples was quantified using the RMS method.

Main Results:

• The RMS method demonstrated robustness and reliability in the quantitative analysis of 5CQA in apple juice across multiple laboratories.
• Using caffeic acid (CA) as a reference compound circumvented the need for a 5CQA analytical standard.
• Results from the RMS method were comparable to those obtained using a traditional calibration curve, validating the RMS approach.

Conclusions:

• The relative molar sensitivity (RMS) method, utilizing caffeic acid (CA) as a reference, is a validated and reliable technique for the quantitative analysis of chlorogenic acid (5CQA).
• This method overcomes the limitations associated with the purity, stability, and hygroscopic nature of 5CQA analytical standards.
• The RMS method provides an accurate alternative for quantifying 5CQA in complex matrices like apple juice without requiring a specific analyte standard.