Danielle Ryan

Sample preparation in the determination of phenolic compounds in fruits

Phenolic compounds occur as secondary metabolites in all plants.1 They embrace a considerable range of substances possessing an aromatic ring bearing one or more hydroxy substituents, although a more precise definition is based on metabolic origin as those substances derived from the shikimate pathway and phenylpropanoid metabolism.2 A convenient classification of the plant phenols distinguishes the number of constitutive carbon atoms in conjunction with the structure of the basic phenolic skeleton (Table 1). The range of known phenolics is thus vast and also includes polymeric lignins and condensed tannins. Some plant phenols may be involved in primary metabolism whereas others have an effect on plant growth or protect the more vulnerable cell constituents against photooxidation by ultraviolet light by virtue of their strong UV absorption.3 Plant phenols also play an important role in disease resistance in the plant. Intense interest in fruit phenolics is also related to their physiological activity which depends on their antioxidant activity, the ability to scavenge both active oxygen species and electrophiles, the ability to inhibit nitrosation and to chelate metal ions, the potential for autooxidation and the capability to modulate certain cellular enzyme activities.4–7 Thus, knowledge of the levels of these compounds in plants is of considerable interest but is limited by problems of analysis. The structural diversity of the phenolics and its effect on physicochemical behaviour such as solubility and analyte recovery presents a challenging analytical problem. Moreover, a number of phenolic compounds are easily hydrolysed and many are relatively easily oxidized, which further complicates sample handling.8,9 This review emphasises the importance of sample preparation in the determination of phenolic compounds in plant materials particularly fruits. Fruits are an important dietary source of phenolic substances although interest is also shifting to other parts of the plant as potential commercial sources of phenols. Sample preparation is a critical step in analysis and this is even more significant with real samples where the matrix components are biologically active and the analytes represent a diverse spectrum of numerous compounds, many having an unknown identity. Thus, methods of extraction of phenols from fruits are generally dependent on several factors while the usual quantification procedures involve the separation sciences and are universally applicable. Soleas et al.10 illustrated this point. They developed a derivatization procedure for determination of 15 phenolic constituents in solid vitaceous plant materials and concluded that the method ‘should be suitable to measure polyphenols in fruit, vegetables, and other foods provided that efficient extraction techniques are employed’. Such statements are seen frequently in the analytical literature but they tend to belittle the importance of this step (or perhaps they serve to underline its critical importance). Rhodes and Price11 observed that the determination of phenolic species in foods is an important outstanding problem and reviewed methods for the extraction and purification of phenolic antioxidants as the conjugated forms that exist in plant foods. Knowledge of the extraction of phenolics is also desirable outside the analytical context for it has important practical applications in the food industry. For instance, the mechanism and kinetics of phenolic extraction from wood to wine during ageing in barrels12 has significant consequences for the production of quality wines.

Biotransformations of phenolic compounds in Olea europaea L.

Phenolic compounds are a diverse range of secondary metabolites derived from the shikimate pathway and phenylpropanoid metabolism. Olea europaea L. contains a number of unusual phenolics including various oleosides. The amounts and types of phenolics vary markedly between leaf, fruit, stone, and seed. The metabolic relationships between the various parts and phenolic content are poorly understood. Interest in this area is related to the importance of the phenolic profile to the aesthetics and quality of olive products, and to the use of olive leaves in phytomedicines.

Recent and potential developments in the analysis of urine: a review.

Analysis of urine is a widely used diagnostic tool that traditionally measured one or, at most, a few metabolites. However, the recognition of the need for a holistic approach to metabolism led to the application of metabolomics to urine for disease diagnostics. This review looks at various aspects of urinalysis including sampling and traditional approaches before reviewing recent developments using metabolomics. Spectrometric approaches are covered briefly since there are already a number of very good reviews on NMR spectroscopy and mass spectrometry and other spectrometries are not as highly developed in their applications to metabolomics. On the other hand, there has been a recent surge in chromatographic applications dedicated to characterising the human urinary metabolome. While developments in the analysis of urine encompassing both classical approaches of urinalysis and metabolomics are covered, it must be emphasized that these approaches are not orthogonal - they both have their uses and are complementary. Regardless, the need to normalise analytical data remains an important impediment.

Determination of phenolic compounds in olives by reversed-phase chromatography and mass spectrometry

Abstract A method based on extraction from freeze-dried material and clean up by solid-phase extraction was optimized for recovery of phenolic compounds from olive fruit. The extracted compounds were characterised by reversed-phase liquid chromatography using both ultraviolet, fluorescence and electrospray ionization mass spectral detection. Using this approach, oleuropein was confirmed as the major phenolic in olive fruit. Other compounds whose presence was confirmed include tyrosol, syringic, ferulic and homovanillic acids, quercetin-3-rhamnoside, ligstroside and isomers of verbascoside. Elenolic acid and its glucoside, which are not phenolic but are closely related, were also identified in sample extracts.

Standardizing the experimental conditions for using urine in NMR-based metabolomic studies with a particular focus on diagnostic studies: a review

AbstractThe metabolic composition of human biofluids can provide important diagnostic and prognostic information. Among the biofluids most commonly analyzed in metabolomic studies, urine appears to be particularly useful. It is abundant, readily available, easily stored and can be collected by simple, noninvasive techniques. Moreover, given its chemical complexity, urine is particularly rich in potential disease biomarkers. This makes it an ideal biofluid for detecting or monitoring disease processes. Among the metabolomic tools available for urine analysis, NMR spectroscopy has proven to be particularly well-suited, because the technique is highly reproducible and requires minimal sample handling. As it permits the identification and quantification of a wide range of compounds, independent of their chemical properties, NMR spectroscopy has been frequently used to detect or discover disease fingerprints and biomarkers in urine. Although protocols for NMR data acquisition and processing have been standardized, no consensus on protocols for urine sample selection, collection, storage and preparation in NMR-based metabolomic studies have been developed. This lack of consensus may be leading to spurious biomarkers being reported and may account for a general lack of reproducibility between laboratories. Here, we review a large number of published studies on NMR-based urine metabolic profiling with the aim of identifying key variables that may affect the results of metabolomics studies. From this survey, we identify a number of issues that require either standardization or careful accounting in experimental design and provide some recommendations for urine collection, sample preparation and data acquisition.

Biosynthesis and biotransformations of phenol-conjugated oleosidic secoiridoids from Olea europaea L.

The genus Olea contains the economically important European olive tree (Olea europaea L.). This species is also of chemotaxonomic interest because of the presence of various phenol-conjugated oleosidic secoiridoids or oleosides. The chemistry of these phenolic oleosides is diverse and complicated, and it is only in recent years that attention has been given to their biosynthesis and the biotransformations during the processing and storage of olive products. Many questions regarding these processes remain unanswered, and yet these have significant impact on the quality and value of olive products such as olive oil.

Liquid chromatography with electrospray ionisation mass spectrometric detection of phenolic compounds from Olea europaea

The results demonstrate the potential of electrospray ionisation mass spectrometry for the specific detection of phenolic species in olives. Phenolic compounds were detected with greater sensitivity in the negative ion mode, but results from positive and negative ion modes were complementary with the positive ion mode showing structurally significant fragments. This is demonstrated by the identification of oleuropein and isomers of verbascoside. The structure of the latter were confirmed by retention, mass spectral and nuclear magnetic resonance data. These isomers have not previously been reported in olive.

Applications of mass spectrometry to plant phenols

Abstract Plant phenols embrace a considerable range of compounds and are defined as those substances derived from the shikimate acid pathway and phenylpropanoid metabolism. The present article examines the application of mass spectrometry to the analysis of these compounds and traces the chronological development of analyte ionisation methods.

Bioactivity of oats as it relates to cardiovascular disease.

The food consumption of oats has increased in recent years due to a perceived association with a range of health benefits. Oats are unusual in that the bran is not as physically distinct as in other cereals. This provides a possible benefit in providing a high beta-glucan content of the grains. However, oats contain many other phytochemicals including a range of antioxidants that may be associated with health benefits, although the evidence for such benefits is largely indirect and often confusing and contradictory. Nevertheless, the consumption of oats as part of a balanced diet does seem a reasonable approach.

Recovery of phenolic compounds from Olea europaea

The recovery of biophenols from olive drupes by simple extraction (free phenolics) was compared to that from acid- and base-treated extracts. Quantitative data are presented for levels of phenols obtained by the different procedures. The major phenol in green olives, oleuropein, was significantly affected by both acid- and base-treatment, and resulted in the liberation of hydroxytyrosol glucoside. The level of hydroxytyrosol glucoside produced after 24 h of acid-treatment was approximately 50 times that produced in base-treated extracts.