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Rapid methods for extracting and quantifying phenolic compounds in citrus rinds.

ABSTRACT: Conventional methods for extracting and quantifying phenolic compounds in citrus rinds are time consuming. Rapid methods for extracting and quantifying phenolic compounds were developed by comparing three extraction solvent combinations (80:20 v/v ethanol:H2O; 70:29.5:0.5 v/v/v methanol:H2O:HCl; and 50:50 v/v dimethyl sulfoxide (DMSO):methanol) for effectiveness. Freeze-dried, rind powder was extracted in an ultrasonic water bath at 35°C for 10, 20, and 30 min. Phenolic compound quantification was done with a high-performance liquid chromatography (HPLC) equipped with diode array detector. Extracting with methanol:H2O:HCl for 30 min resulted in the optimum yield of targeted phenolic acids. Seven phenolic acids and three flavanone glycosides (FGs) were quantified. The dominant phenolic compound was hesperidin, with concentrations ranging from 7500 to 32,000 μg/g DW. The highest yield of FGs was observed in samples extracted, using DMSO:methanol for 10 min. Compared to other extraction methods, methanol:H2O:HCl was efficient in optimum extraction of phenolic acids. The limit of detection and quantification for all analytes were small, ranging from 1.35 to 5.02 and 4.51 to 16.72 μg/g DW, respectively, demonstrating HPLC quantification method sensitivity. The extraction and quantification methods developed in this study are faster and more efficient. Where speed and effectiveness are required, these methods are recommended.

SUBMITTER: Magwaza LS

PROVIDER: S-EPMC4708637 | biostudies-other | 2016 Jan

REPOSITORIES: biostudies-other

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Project description:Background and aimsPhenolic compounds contribute to food quality and have potential health benefits. Consequently, they are an important target of selection for Citrus species. Numerous studies on this subject have revealed new molecules, potential biosynthetic pathways and linkage between species. Although polyphenol profiles are correlated with gene expression, which is responsive to developmental and environmental cues, these factors are not monitored in most studies. A better understanding of the biosynthetic pathway and its regulation requires more information about environmental conditions, tissue specificity and connections between competing sub-pathways. This study proposes a rapid method, from sampling to analysis, that allows the quantitation of multiclass phenolic compounds across contrasting tissues and cultivars.MethodsLeaves and fruits of 11 cultivated citrus of commercial interest were collected from adult trees grown in an experimental orchard. Sixty-four phenolic compounds were simultaneously quantified by ultra-high-performance liquid chromatography coupled with mass spectrometry.Key resultsCombining data from vegetative tissues with data from fruit tissues improved cultivar classification based on polyphenols. The analysis of metabolite distribution highlighted the massive accumulation of specific phenolic compounds in leaves and the external part of the fruit pericarp, which reflects their involvement in plant defence. The overview of the biosynthetic pathway obtained confirmed some regulatory steps, for example those catalysed by rhamnosyltransferases. The results suggest that three other steps are responsible for the different metabolite profiles in 'Clementine' and 'Star Ruby' grapefruit.ConclusionsThe method described provides a high-throughput method to study the distribution of phenolic compounds across contrasting tissues and cultivars in Citrus, and offers the opportunity to investigate their regulation and physiological roles. The method was validated in four different tissues and allowed the identification and quantitation of 64 phenolic compounds in 20 min, which represents an improvement over existing methods of analysing multiclass polyphenols.

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Rapid methods for extracting and quantifying phenolic compounds in citrus rinds.

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