A New UHPLC-UV-MS Method for Detection and Identification of Bioactive Compounds.

Abstract

Background: The development of analytical methods in food science has grown parallel with consumers' concern about food composition. There is desire to better understand the phytochemicals present in foods and feeds. Comprehensive screening of nutrients and bioactive compounds can be challenging due to the vast differences in molecular properties of the compounds.

Objective: To develop a sensitive liquid chromatography-mass spectrometry (LC-MS) analytical method to detect, identify, and quantify hundreds of compounds from a broad range of molecular categories in a single method, from hydrophilic (e.g., organic acids) to hydrophobic (e.g., sterols) nutrients.

Method: The chromatographic separation was tailored for a balanced detection of hydrophilic and hydrophobic compounds, with attention towards the separation of isomeric polyphenols. The chromatographic separation was performed in 45 min on a C18-PFP stationary phase, which provided enhanced resolution of isomers. The method employs two columns in series to improve the resolution, which requires ultra-high-pressure instrumentation (1,000 bar). The addition of an in-line ultraviolet (UV) diode-array detector allowed for spectral profile confirmation of the identities of many targeted and unknown compounds.

Results: The developed method was applied in this work to measure different types of bioactives recognized to have health benefits: flavonoids, phenolic acids, vitamins, tocochromanols, phytosterols, sugars, organic acids, lipids, amines, nucleic acids, and many other small molecules. Four chocolate products were analyzed as a demonstration of the range of the method.

Conclusions: This method has enabled discoveries of new commercial value in the food processing industry by capturing information on a wider range of analytes than previous individual methods, and this data can be leveraged to promote the health and well-being of consumers.

Highlights: This new LC-UV-MS methodology provides a more comprehensive analysis of a broad range of molecularly diverse bioactive constituents in raw materials and finished products in the food and feed industry.