An integrated analytical approach based on NMR, LC-MS and GC-MS to evaluate thermal and non-thermal processing of cashew apple juice.

Innovative chemometric approaches by NMR and LC-MS data fusion (multiblock analysis) and decomposition of the GC-MS raw data by PARADISe were applied to evaluate the influence of thermal and non-thermal processing on the composition of cashew apple juices. Comparative investigations by Principal Component Analysis (PCA) highlighted pronounced effect of thermal processing on juice compared to non-thermal processing, as decreases of anacardic acids, sucrose, malic acid, tyrosine, phenylalanine, and important flavor compounds (esters, aldehydes, and ketones). Ultrasound presented relevant influence on increase of anacardic acids concentration. Non-thermal processing carried out at more intense conditions (10 min of ultrasound, 5 min of ozone, and pulsed light at 10 V) showed pronounced effect compared to other non-thermal processing. Although individual PCA enables to detect the influence of different processing technologies, data fusion and PARADISe presented advantages, since a more comprehensive understanding of the relationship among chemical changes from different analytical techniques were established.

Cold plasma processing effect on cashew nuts composition and allergenicity.

The study investigated the influence of atmospheric plasma processing on cashew nut composition as well as on its allergenicity. The cashew nuts were processed by low-pressure plasma, using glow discharge plasma (80 W and 50 kHz power supply). Anacardic acids and allergens were quantified by HPLC and immunoassay, respectively. Additionally, the overall composition was evaluated by 1H qNMR. Increases in amounts of anacardic acids (15:1, 15:2, and 15:3) and fatty acids (oleic, linoleic, palmitic and stearic) were detected after all process conditions, with 70.92% of total variance captured using 2 LVs. The total amount of anacardic acids increased from 0.7 to 1.2 µg·mg-1 of nut. The major change was observed for anacardic acid (C15:3) with an increase from 0.2 to 0.55 µg/mg of nut for the samples treated with a flow of 10 mL·min-1 and 30 min of processing. On the other hand, the amount of sucrose decreased, from 33 to 18 mg·g-1 of nut, after all processing conditions. Plasma processing of cashew nuts did not affect binding of either the rabbit anti-cashew or human cashew allergic IgE binding. Among the treatments, 10 min of plasma processing at flow rate of 30 mL·min-1 of synthetic air followed by 20 min at flow rate 5.8 mL·min-1 had the least effect on nut composition as a whole.

Photoelectrochemical determination of the activity of histone acetyltransferase and inhibitor screening by using MoS2 nanosheets.

The enzyme histone acetyltransferase (HAT) catalyzes the acetylation of a substrate peptide, and acetyl coenzyme A is converted to coenzyme A (CoA). A photoelectrochemical method is described for the determination of the HAT activity by using exfoliated MoS2 nanosheets, phos-tag-biotin, and ß-galactosidase (ß-Gal) based signal amplification. The MoS2 nanosheets are employed as the photoactive material, graphene nanosheets as electron transfer promoter, gold nanoparticles as recognition and capture reagent for CoA, and phos-tag-biotin as the reagent to link CoA and ß-Gal. The enzyme ß-Gal catalyzes the hydrolysis of substrate O-galactosyl-4-aminophenol to generate free 4-aminophenol which is a photoelectrochemical electron donor. The photocurrent increases with the activity of HAT. Under optimal conditions, the response is linear in the 0.3 to 100 nM activity range, and the detection limit is 0.14 nM (at S/N = 3). The assay was applied to HAT inhibitor screening, specifically for the inhibitors C646 and anacardic acid. The IC50 values are 0.28 and 39 µM, respectively. The method is deemed to be a promising tool for epigenetic research and HAT-targeted cancer drug discovery. Graphical abstract Histone acetyltransferase was detected using a sensitive photoelectrochemical method using MoS2 nanosheets as photoactive material.

Application of multivariate optimization for the selective extraction of phenolic compounds in cashew nuts (Anacardium occidentale L.).

An extraction method based on a multivariate analytical approach was developed for enhancement of the phenolic compounds in cashew nut extracts. The different extractor solvents (acetone, water, ethanol, and methanol) and their binary, ternary, and quaternary combinations were evaluated using a simplex-centroid design and surface response methodology. The special cubic model exhibits no lack of fit and explains 89.2% of the variance. The total phenolic measurements by the Folin-Ciocalteu method revealed the highest values for ethanol (5.93 mg GAE g-1) and acetone-methanol-ethanol ternary mixture (5.92 mg GAE g-1) extracts. ESI (-)-Q/TOFMS analyses combined with PCA and HCA revealed the presence of fatty acids, phospholipids, and sugars in the ternary mixture cashew extract, while for the ethanol extract only phenolic compounds, such as anacardic acids and derivatives, were found. The proposed approach was adequate to reach the optimal extractor which ethanol, a low-toxicity solvent, enabled the selective extraction of a high content of phenolic compounds from cashew nuts.

A highly sensitive GC-MS method for simultaneous determination of anacardic acids in cashew (Anacardium occidentale) nut shell oil in the presence of other phenolic lipid derivatives.

The commercial value of cashew nut shell liquid (CNSL) has become a cornerstone of the agrowaste industry. It is the by-product of the cashew industry and has an 1/8 inch thickness of soft honeycomb structure. CNSL contains phenolic lipids with aliphatic chains such as anacardic acid, cardanol, cardol and methyl cardol, and their derivatives. The developed GC-MS method is rapid, accurate and selective using a selected derivatizing reagent, namely N-methyl-N-(trimethylsilyl)-trifluoroacetamide that was previously diluted 1:1% with anhydrous pyridine. The proposed GC-MS method was applied for the analysis of different CNSL samples. The results showed that all classes of CNSL compounds were detected. The four alkyl phenols were detected with their different alkyl sidechains without any interference. This method is also specified for the detection of fatty acids of saturated and unsaturated chains. Silylation did not cause any alteration in the chemical structure of CNSL compounds regardless of esterification action. Silylation is considered a safe derivatizing agent compatible with GC chromatography and specific for all volatile and nonvolatile polar and nonpolar CNSL compounds that could be detected in CNSL samples.

Development and Validation of a Reversed Phase HPLC Method for Determination of Anacardic Acids in Cashew (Anacardium occidentale) Nut Shell Liquid.

Cashew nut shell liquid (CNSL) contains phenolic lipids with aliphatic chains that are of commercial interest. In this work, a chromatographic method was developed to monitor and quantify anacardic acids (AnAc) in CNSL. Samples containing AnAc were analyzed on a high-performance liquid chromatograph coupled to a diode array detector, equipped with a reversed phase C18 (150 × 4.6 mm × 5 µm) column using acetonitrile and water as the mobile phase both acidified with acetic acid to pH 3.0 in an isocratic mode (80:20:1). The chromatographic method showed adequate selectivity, as it could clearly separate the different AnAc. To validate this method, AnAc triene was used as an external standard at seven different concentrations varying from 50 to 1,000 µg mL-1. The Student's t-test and F-test were applied to ensure high confidence for the obtained data from the analytical calibration curve. The results were satisfactory with respect to intra-day (relative standard deviation (RSD) = 0.60%) and inter-day (RSD = 0.67%) precision, linearity (y = 2,670.8x - 26,949, r2 > 0.9998), system suitability for retention time (RSD = 1.02%), area under the curve (RSD = 0.24%), selectivity and limits of detection (19.8 µg mg-1) and quantification (60.2 µg mg-1). The developed chromatographic method was applied for the analysis of different CNSL samples, and it was deemed suitable for the quantification of AnAc.

UPLC-qTOF-MS/MS-based phenolic profile and their biosynthetic enzyme activity used to discriminate between cashew apple (Anacardium occidentale L.) maturation stages.

Cashew immature and ripe peduncles (Anacardium occidentale L.) from orange- and red-colored clones CCP 76 and BRS 189, respectively, were prepared as juice or fibrous fraction and submitted to UPLC-MS analyses, while the soluble fraction was also submitted to enzymatic evaluation. Cinnamoyl glucoside was present in ripe juice samples from both cashew clones, while monogalloyl diglucoside and digalloyl glucoside were present in immature juice samples from both cashew clones. Four compounds were found at immature fiber of both clones, anacardic acids (1, 2, 3) and GA19. The phenolic biosynthetic pathway was evaluated in juice samples and phenylalanine ammonia-lyase activity decreased significantly during the development, although it was much higher in ripe CCP 76. UDP-glycosyltransferases activity differed between clones, however its product cinnamoyl glucoside was a possible chemical marker of ripe juice samples from both clones. Flavonol synthase showed the highest specific activity in both cashew clones and its product, flavonols were identified in cashew apple at immature and ripe stages.

Effects of immature cashew nut-shell liquid (Anacardium occidentale) against oxidative damage in Saccharomyces cerevisiae and inhibition of acetylcholinesterase activity.

The cashew tree (Anacardium occidentale) represents one of the major cheapest sources of non-isoprenoid phenolic lipids, which have a variety of biological properties: they can act as molluscicides, insecticides, fungicides, have anti-termite properties, have medicinal applications, and demonstrate antioxidant activity in vitro. Immature cashew nut-shell liquid (iCNSL) is a unique natural source of unsaturated long-chain phenols. Their use has stimulated much research in order to prepare drug analogues for application in several fields. The objective of the present study was to determine whether iCNSL has antioxidant properties when used in strains of the yeast Saccharomyces cerevisiae and to measure the inhibitory activity of acetylcholinesterase. The constituents were identified using thin-layer chromatography, gas chromatography-mass spectrometry, Fourier transform infrared spectroscopy, and (1)H and (13)C nuclear magnetic resonance. The iCNSL contains anacardic acid, cardanol, cardol, and 2-methyl cardol. Immature cashew nut oil contains triacylglycerols, fatty acids, alkyl-substituted phenols, and cholesterol. The main constituents of the free fatty acids are palmitic (C(16:0)) and oleic acid (C(18:1)). iCNSL has excellent protective activities in strains of S. cerevisiae against oxidative damage induced by hydrogen peroxide and inhibits acetylcholinesterase activity. iCNSL may have an important role in protecting DNA against damage induced by reactive oxygen species, as well as hydrogen peroxide, generated by intra- and extracellular mechanisms.

Characterization of anacardic acids by micellar electrokinetic chromatography and mass spectrometry.

A possibility of using capillary electrophoresis for separation of anacardic acids (6-alkylsalicylic acids) has been studied. Conventional micellar electrokinetic chromatography (MEKC) in non-coated fused silica capillaries and reversed-flow micellar electrokinetic chromatography (RF-MEKC) in capillaries coated with polydimethylacrylamide was applied for separation of anacardic acids extracted from cashew nuts. Influence of the composition of background electrolyte on the resolution of anacardic acid isomers was evaluated. Separations were performed using sodium dodecyl sulphate (SDS) micelles and mixed micelles of SDS and polyoxyethylene lauryl ether as a pseudostationary phase. To further improve the separation in RF-MEKC, beta-cyclodextrin and a dual cyclodextrin system of beta-cyclodextrin with heptakis-6-sulphato-beta-cyclodextrin was added to the working electrolyte. Best separation of anacardic acids were achieved in the polydimethylacrylamide-coated capillary using 10 mM phosphate background electrolyte pH 6.5 with addition of 1 M urea, 20% acetonitrile, 10 mM of beta-cyclodextrin and 1 mM of heptakis-6-sulfo-beta-cyclodextrin. Mass spectrometry was used for the identification of anacardic acids in the extract from cashew nuts in single and tandem mode using Q-TOF instrument. Nine anacardic acids were identified in the extract form the cashew nuts.

6-alkylsalicylic acids and 6-alkylresorcylic acids from ants in the genus Crematogaster from Brunei.

The defensive chemistry of two species of ants from Brunei in the genus Crematogaster (Physocrema group) has been investigated. Ants in this group release a white secretion from hypertrophied metapleural glands on their thorax when they are disturbed. Previously, one species in this group has been shown to produce alkylphenols and alkylresorcinols. In the present investigation, similar compounds along with salicylic acids and resorcylic acids that are anacardic acid and olivetolic acid homologs, respectively, are described from two species. The structures of these compounds were suggested by their spectroscopic data and confirmed by direct comparison with synthetic samples. Some of these compounds occur in lichens and have well documented physiological activities.