Identification and quantification of cyclitols and sugars isolated from different morphological parts of Raphanus sativus L.

Through this study, we aimed to develop a new analytical method for identification and quantification of sugars and cyclitols isolated from different morphological parts of Raphanus sativus L (R. sativus). Accelerated solvent extraction with water was involved for targets extraction. Solid phase extraction was used for purification and preconcentration, while high performance liquid chromatography with evaporative light scattering detector (HPLC-ELSD) was used for chromatographic analyses. A short method of only 30 min for a single analysis was developed finally. The obtained results, allowed for quantification of eight targets, i.e., three cyclitols (D-pinitol, allo-inositol and scyllo-inositol) and five sugars (xylose, D-mannose, D-fructose, D-glucose and sucrose) that were determined simultaneously using a single analysis. The developed method can be applied in industry as a routine method for analysis of sugars and cyclitols from different sources.

New Methodology for the Identification of Metabolites of Saccharides and Cyclitols by Off-Line EC-MALDI-TOF-MS.

A combination of electrochemistry (EC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (off-line EC-MALDI-TOF-MS) was applied for determination of the studied biologically active compounds (D-glucose, D-fructose, D-galactose, D-pinitol, L-chiro-inositol, and myo-inositol) and their possible electrochemical metabolites. In this work, boron-doped diamond electrode (BDD) was used as a working electrode. MALDI-TOF-MS experiments were carried out (both in positive and negative ion modes and using two matrices) to identify the structures of electrochemical products. This was one of the first applications of the EC system for the generation of electrochemical products produced from saccharides and cyclitols. Moreover, exploratory data analysis approaches (correlation networks, hierarchical cluster analysis, weighted plots) were used in order to present differences/similarities between the obtained spectra, regarding the class of analyzed compounds, ionization modes, and used matrices. This work presents the investigation and comparison of fragmentation patterns of sugars, cyclitols, and their respective products generated through the electrochemistry (EC) process.

Correlation Study of Honey Regarding their Physicochemical Properties and Sugars and Cyclitols Content.

Honey is a natural sweetener, with an osmotic effect on microorganisms due to the increased sugar content and low amount of water. Cyclitols are minor constituents of honey. They play a defensive role in plants against unfavorable environmental conditions. Honey's physicochemical properties can vary, resulting in a wide range of colors, flavors, scents, antioxidant activity, dissimilar values of pH, acidity, electrical conductivity, etc. Some literature regarding correlation between honey types is already available, but a comprehensive study displaying an ample evaluation of multifarious aspects is still needed. This study focuses on the correlation between 18 honey types, originating from 10 countries, collected during four years, summarizing a total of 38 samples. A total of 6 physicochemical properties and 18 target components (sugars and cyclitols) were considered as variables. A correlation analysis is presented between the investigated parameters and between honey types, together with the statistical analysis which allowed for observation of the clusters' distribution according with the investigated variables.

Forced degradation of l-(+)-bornesitol, a bioactive marker of Hancornia speciosa: Development and validation of stability indicating UHPLC-MS method and effect of degraded products on ACE inhibition.

The antihypertensive activity of the medicinal plant Hancornia speciosa has been previously demonstrated by us, being the activity ascribed to polyphenols and cyclitols like l-(+)-bornesitol. We herein evaluated the stability of the bioactive marker bornesitol submitted to forced degradation conditions. Bornesitol employed in the study was isolated from H. speciosa leaves. An UHPLC-ESI-MS/MS method was developed to investigate bornesitol stability based on MRM (Multiple Reaction Monitoring) acquisition mode and negative ionization mode, employing both specific (m/z 193 → 161 Da) and confirmatory (m/z 193 → 175 Da) transitions. A gradient elution of 0.1% formic acid in water and acetonitrile was performed on a HILIC column. The method was validated and showed adequate linearity (r2 > 0.99), selectivity, specificity, accuracy, and precision (RSD < 2.9%). The method was robust for deliberate variations on dessolvation temperature, but not for changes in the flow rate and dessolvation gas. The results from the stability studies allowed us to classify bornesitol as labile for acidic and alkaline hydrolysis, but as very stable for oxidative and neutral hydrolysis exposure. Bornesitol was categorized as practically stable under photolysis degradation, whereas a considerable reduction on its contents was induced by metal ions and thermolysis exposure. Degraded samples from neutral hydrolysis and thermolysis were assayed in vitro for ACE inhibition and showed a substantial decrease in biological activity as compared to intact bornesitol. myo-Inositol was identified as the major degradation products in both matrices. This is the first report on bornesitol stability under different stress conditions and the obtained data are relevant for the development and quality control of standardized products from H. speciosa leaves.

New approach for fast identification of cyclitols by MALDI-TOF mass spectrometry.

INTRODUCTION: Alfalfa (Medicago sativa L.) is the subject of many studies due to its numerous chemical constituents and beneficial properties. Among these constituents are cyclitols, which have attracted attention due to the variety of biological properties they have. OBJECTIVE: A rapid and sensitive analytical procedure based on matrix-assisted laser desorption ionisation technique with time-of-flight and mass spectrometry (MALDI-TOF-MS) analysis was used for the first time for the identification of three cyclitols from different parts of alfalfa. METHODOLOGY: Plant extracts were prepared and purified using Soxhlet extraction and solid-phase extraction (SPE). Then, samples were dissolved in α-cyano-4-hydroxycinnamic acid (HCCA) matrix, and subjected to MALDI-TOF-MS analysis. RESULTS: The ion at m/z 524.0 was distributed in all standards and in leaves and stem extracts. In turn, the signal at m/z 335.1 was found in all standards and all alfalfa extracts. The ion at m/z144.1 was found just for d-chiro-inositol and distributed in all extracts. Both signals at m/z 265.9 and 250.0 were found only in l-chiro-inositol standard and the extract of stem. However, the ion at m/z 177.1 was found in d-pinitol standard and the extract of leaves. Based on molecular weights, information on fragment ions obtained by MALDI-TOF-MS, and the chemistry of cyclitols, we successfully identified three cyclitols (d-chiro-inositol, l-chiro-inositol, d-pinitol) in different parts of alfalfa (leaves, stem, flowers). CONCLUSION: The obtained results in this study proved that MALDI-TOF-MS is a rapid, sensitive and very powerful tool for identification of cyclitols within plants and has the potential to differentiate between enantiomers.

Determination of sugars and cyclitols isolated from various morphological parts of Medicago sativa L.

Plant research interest has increased all over the world, and a large body of evidence has been collected to show the huge potential of medicinal plants in various disease treatments. Medicago sativa L., known as alfalfa, is a rich source of biologically active components and secondary metabolites and was frequently used from the ancient times both as fodder crop and as a traditional medicine in the treatment of various diseases. Cyclitols, naturally occurring in this plant, have a particular interest for us due to their significant anti-diabetic, antioxidant, anti-inflammatory, and anti-cancer properties. In the present study we revealed the isolation, the identification, and the quantification of some cyclitols and sugars extracted from different morphological parts of alfalfa plant. Soxhlet extraction and solid phase extraction were used as extraction and purification methods, while for the analyses derivatization followed by gas chromatography with mass spectrometry was involved. The obtained results showed significant differences in the quantities of cyclitols and sugars found in the investigated morphological parts, ranging between 0.02 and 13.86 mg/g of plant in case of cyclitols, and in the range of 0.09 and 40.09 mg/g of plant for sugars. However, roots have the richest part of cyclitols and sugars in contrast to the leaves.

Development and validation of an HPLC-DAD method for quantification of bornesitol in extracts from Hancornia speciosa leaves after derivatization with p-toluenesulfonyl chloride.

Cylitol L-(+)-bornesitol is regarded as a bioactive constituent from Hancornia speciosa leaves, a plant species traditionally used in Brazil to treat diabetes and hypertension. We report a simple HPLC-DAD method for the quantification of bornesitol in extracts from H. speciosa leaves, after derivatization with p-toluenesulfonyl chloride, using pentaerythritol as internal standard. A gradient of methanol, acetonitrile and water was employed for elution on an ODS column and detection was set up at a wavelength of 230 nm. The method was selective and linear over the range 60.4-302.0 µg/ml with r² of 0.9981, and showed satisfactory precision for intra-day (RSD=2.37%) and inter-day (RSD=3.17%) assays. The recovery varied between 92.3% and 99.9% and the limits of quantification and detection were respectively 5.00 and 1.67 µg/ml. The method was applied to quantify bornesitol in extracts from H. speciosa leaves of different specimens.

Strength from weakness: opportunistic CH···O hydrogen bonds differentially dictate the conformational fate in solid and solution states.

We show that the CH···O hydrogen bond can be opportunistic to differentially dictate the conformation of cyclitols in solution and solid states. While several intermolecular CH···O stabilize the chair-conformation in the solid, single intramolecular CH···O stabilizes an otherwise unfavorable boat-conformation in solutions analogous to the boat-conformation of cis-1,4-cyclohexanediols by OH···O bonds.

Free cyclitol, soluble carbohydrate and protein contents in Vigna unguiculata and Phaseolus vulgaris bean sprouts.

Seeds sprouts have been used as a good source of basic nutrients and nutraceutical compounds. The high nutritional value of seeds derives from the deposition of compounds during development. However some of these molecules are used in metabolic processes like germination, which leads to a considerable variation in their concentrations once these events are completed. In this work, we investigate the levels of inositols (myo-inositol, D-pinitol and ononitol), soluble carbohydrates and proteins in cotyledons of Phaseolus vulgaris and Vigna unguiculata sprouts. Sprouting increased myo-inositol and glucose content and reduction of raffinose and ononitol was observed. The protein levels increased in P. vulgaris and decreased in V. unguiculata sprouting. The level of sucrose was maintained in both sprouts. D-Pinitol was detected only in quiescent seeds. Our results suggested that bean sprout is an important source of proteins, sucrose, glucose and myo-inositol. Additionally, bean sprouts have low levels of raffinose, an antinutritional compound.

Cyclic polyalcohols: fingerprints to identify the botanical origin of natural woods used in wine aging.

Cyclic polyalcohol composition of 80 natural wood samples from different botanical species, with the majority of them used in the oenology industry for aging purposes, has been studied by gas chromatography-mass spectrometry (GC-MS) after its conversion into their trimethylsilyloxime derivatives. Each botanical species showed a different and specific cyclic polyalcohol profile. Oak wood samples were characterized by the richness in deoxyinositols, especially proto-quercitol. Meanwhile, other botanical species showed a very low content of cyclic polyalcohols. The qualitative and quantitative study of cyclic polyalcohols was a useful tool to characterize and differentiate woods of different botanical origin to guarantee the authenticity of chips used in the wine-aging process. Monosaccharide composition was also analyzed, showing some quantitative differences among species, but cyclic polyalcohols were the compounds that revealed the main differentiation power.

Analysis of cyclitols in different Quercus species by gas chromatography-mass spectrometry.

BACKGROUND: The carbohydrate profile of some woods used for aging wines and spirits has been recently studied using a pressurized liquid extraction method, the main differences found being related to cyclitol content. The aim of this study was to perform a detailed study of these compounds in woods of different Quercus species in order to identify two unknown compounds which appeared in the extracts and to verify whether the obtained profile was homogeneous for other Quercus species. RESULTS: Besides the known monosaccharides and five cyclitols previously described, three deoxy-inositols (epi-, vibo- and scyllo-quercitol) were identified. The presence of these eight cyclitols was confirmed in all subgenera and species of Quercus analyzed, allowing a characteristic cyclitol profile. CONCLUSIONS: Three deoxy-inositols (quercitols) have been identified in the carbohydrate profile of oak wood. All examined Quercus species displayed a common profile consisting of four inositols and four quercitols, which represent a good dataset for characterization of this genus.