[Optimization for determination methods of alpha-, beta- carotene in vegetables]

Area of Science:

• Analytical Chemistry
• Food Science
• Nutritional Biochemistry

Background:

• Carotenoids, including alpha-carotene and beta-carotene, are essential plant pigments with significant health benefits.
• Accurate quantification of individual carotenes is crucial for nutritional analysis and food quality assessment.
• Existing methods may lack the resolution to effectively separate carotene isomers.

Purpose of the Study:

• To develop and optimize a high-performance liquid chromatography (HPLC) method for the precise separation and determination of alpha-carotene and beta-carotene.
• To validate the optimized method for its accuracy, reliability, and suitability for analyzing fresh vegetables.

Main Methods:

• Sample preparation involved ethanol extraction, saponification, petroleum ether extraction, washing, drying, and concentration.
• Separation was achieved using a YMC Carotenoid C30 column (3 µm x 4.6 mm x 150 mm) with a linear gradient elution.
• The mobile phase consisted of methanol:acetonitrile:water and methyl tert-butyl ether, with detection at 450 nm.

Main Results:

• The C30-HPLC column demonstrated excellent separation of carotenes and their isomers.
• Optimal extraction involved heating with anhydrous ethanol before saponification.
• The method showed high linearity (R² > 0.999), good precision (RSD < 10%), and recovery rates (81.6%-93.3%) for both alpha- and beta-carotene.

Conclusions:

• The optimized HPLC method is accurate, reliable, and suitable for the quantitative analysis of alpha-carotene and beta-carotene in fresh vegetables.
• This validated method facilitates precise carotene profiling in food matrices.
• The findings contribute to improved analytical techniques in food and nutritional science.