A rapid validated UHPLC-PDA method for anthocyanins quantification from Euterpe oleracea fruits

Area of Science:

• Analytical Chemistry
• Food Science
• Pharmacognosy

Background:

• Euterpe oleracea (açai) is rich in anthocyanins, potent antioxidants with potential health benefits.
• Accurate quantification of these compounds is crucial for quality control and understanding their bioavailability.
• Existing methods for anthocyanin analysis in açai may lack efficiency or comprehensive validation.

Purpose of the Study:

• To develop and validate the first Ultra-High-Performance Liquid Chromatography-Photodiode Array (UHPLC-PDA) method for major anthocyanins in Euterpe oleracea.
• To establish fast extraction and sample preparation procedures for improved analytical throughput.
• To ensure the reliability and accuracy of the developed method for anthocyanin quantification.

Main Methods:

• Development of a UHPLC-PDA method utilizing an HSS C18 column with gradient elution (acetonitrile and 5% formic acid).
• Implementation of fast extraction procedures involving sequential ethyl acetate and methanol washes.
• Validation of the method using total error and accuracy profiles, including matrix calibration, trueness, repeatability, intermediate precision, selectivity, and linearity.

Main Results:

• A rapid UHPLC-PDA method with a total run time of 17 minutes was successfully developed and validated.
• Matrix calibration proved more accurate than calibration without matrix for anthocyanin quantification.
• The method demonstrated excellent performance with trueness (<6.76% relative bias) and repeatability (<4.6% RSD) for cyanidin-3-glucoside and cyanidin-3-rutinoside within a 1-48 μg/mL range.
• Two novel cyanidin-di-O-glycosides were detected in Euterpe oleracea for the first time.

Conclusions:

• The developed UHPLC-PDA method is the first validated approach for quantifying major anthocyanins in Euterpe oleracea after optimized extraction.
• The sequential extraction strategy effectively removes interfering lipophilic compounds, enhancing anthocyanin analysis.
• This validated method provides a reliable tool for accurate anthocyanin profiling in Euterpe oleracea, supporting research and quality assessment.