Optimization of SPME-Arrow-GC/MS Method for Determination of Free and Bound Volatile Organic Compounds from Grape Skins.

(1) Background: Solid phase microextraction (SPME)-Arrow is a new extraction technology recently employed in the analysis of volatiles in food materials. Grape volatile organic compounds (VOC) have a crucial role in the winemaking industry due to their sensory characteristics of wine.; (2) Methods: Box-Behnken experimental design and response surface methodology were used to optimise SPME-Arrow conditions (extraction temperature, incubation time, exposure time, desorption time). Analyzed VOCs were free VOCs directly from grape skins and bound VOCs released from grape skins by acid hydrolysis.; (3) Results: The most significant factors were extraction temperature and exposure time for both free and bound VOCs. For both factors, an increase in their values positively affected the extraction efficiency for almost all classes of VOCs. For free VOCs, the optimum extraction conditions are: extraction temperature 60 °C, incubation time 20 min, exposure time 49 min, and desorption time 7 min, while for the bound VOCs are: extraction temperature 60 °C, incubation time 20 min, exposure time 60 min, desorption time 7 min.; (4) Conclusions: Application of the optimized method provides a powerful tool in the analysis of major classes of volatile organic compounds from grape skins, which can be applied to a large number of samples.

The Concentration of Organic Acids in Cranberry Juice Modulates the Gut Microbiota in Mice.

A daily consumption of cranberry juice (CJ) is linked to many beneficial health effects due to its richness in polyphenols but could also awake some intestinal discomforts due to its organic acid content and possibly lead to intestinal inflammation. Additionally, the impact of such a juice on the gut microbiota is still unknown. Thus, this study aimed to determine the impacts of a daily consumption of CJ and its successive deacidification on the intestinal inflammation and on the gut microbiota in mice. Four deacidified CJs (DCJs) (deacidification rates of 0, 40, 60, and 80%) were produced by electrodialysis with bipolar membrane (EDBM) and administered to C57BL/6J mice for four weeks, while the diet (CHOW) and the water were ad libitum. Different parameters were measured to determine intestinal inflammation when the gut microbiota was profiled. Treatment with a 0% DCJ did not induce intestinal inflammation but increased the gut microbiota diversity and induced a modulation of its functions in comparison with control (water). The effect of the removal of the organic acid content of CJ on the decrease of intestinal inflammation could not be observed. However, deacidification by EDBM of CJ induced an additional increase, in comparison with a 0% DCJ, in the Lachnospiraceae family which have beneficial effects and functions associated with protection of the intestine: the lower the organic acid content, the more bacteria of the Lachnospiraceae family and functions having a positive impact on the gut microbiota.

Progress in pretreatment and analysis of organic Acids: An update since 2010.

Organic acids, as an important component of food, have great influence on the flavor, texture, freshness of food. By lowering the pH of food to bacteriostatic acidity, organic acids are also used as additives and preservatives. Because organic acids are crucial to predict and evaluate food maturity, production and quality control, the rapid and sensitive determination methods of organic acids are necessary. This review aims to summarize and update the progress of the determination of organic acids in food samples. Pretreatment methods include simple steps (e.g., "dilute and shoot," protein precipitation, filtration, and centrifugation) and advanced microextraction methods (e.g., hollow fiber liquid phase microextraction, stir bar sorptive extraction and dispersive micro-solid phase extraction). Advances in novel materials (nanomaterial), solvents (ionic liquids and supercritical fluids) and hybrid methods are clearly displayed in detail. Continuous progress which has been made in electrochemical method, two-dimensional chromatography, high resolution mass is thoroughly illustrated.

Green microfluidic liquid-phase microextraction of polar and non-polar acids from urine.

In this work, hippuric acid (log P = 0.5), anthranilic acid (log P = 1.3), ketoprofen (log P = 3.6), and naproxen (log P = 3.0) were simultaneously extracted by a green microfluidic device based on the principles of liquid-phase microextraction (LPME). Different deep eutectic solvents (DESs) were investigated as supported liquid membrane (SLM), and a mixture of camphor and menthol as eutectic solvents in the molar ratio 1:1 was found to be highly efficient for the simultaneous extraction of non-polar and polar acidic drugs. LPME was conducted for 6 min per sample. Urine sample was delivered to the system at 1 µL min-1, and target analytes were extracted exhaustively (75-100% recovery) across the DES SLM, and into pure aqueous phosphate buffer pH 11.0 delivered as acceptor at 1 µL min-1. The acceptor was analyzed with liquid chromatography-UV detection. Interestingly, the DES enabled extraction of both the polar and non-polar model analytes at the same time; all chemicals were green and non-hazardous, and the chemical waste was less than 1 mg per sample.

Quantification of ustalic acid, a chemotaxonomic marker, in Tricholoma ustale using liquid chromatography-mass spectrometry.

The development of methods for the detection and qualification of toxic substances in mushrooms is a rapidly growing research area in forensic toxicology. This study aimed to determine liquid chromatographic and mass spectrometric conditions applicable to the analysis of ustalic acid (UA) in Tricholoma ustale. We used HPLC coupled with single quadrupole mass spectrometry (Q-MS) with electrospray ionization in its negative ion mode to analyze UA. We performed HPLC separations on a C18 reversed-phase column with gradient elution using mobile phases containing water, acetonitrile, and formic acid. The MS showed that UA formed the deprotonated molecular ion [M-H]- at m/z 337, which was sufficient for the quantitative analysis of the compound. The average recovery rates of UA from four edible mushrooms (Flammulina velutipes, Pleurotus eryngii, Lentinula edodes, and Grifola frondosa) to which 10.0 µg/g of UA was added were 108%, 104%, 108%, and 107%, respectively, and the relative standard deviation values ranged from 4.1 to 6.4%. Quantitative analysis of UA in three systematically collected individual mushrooms of T. ustale revealed 41.9-155.7 ppm in each dry material. We also explored the fragmentation behaviors of UA in triple quadrupole mass spectrometry and the proposed structures for the product ions. The data suggest that conventional Q-MS with authenticated UA would be able to identify this compound in T. ustale when used for the immediate inspection of incidences of poisoning. Confirmation of the presence of UA in T. ustale would ultimately allow for the chemotaxonomic discrimination of Tricholoma species.

Method development and validation for the quantification of organic acids in microbial samples using anionic exchange solid-phase extraction and gas chromatography-mass spectrometry.

Organic acids play a key role in central metabolic functions of organisms, are crucial for understanding regulatory processes and are ubiquitous inside the cell. Therefore, quantification of these compounds provides a valuable approach for studying dynamics of metabolic processes, in particular when the organism faces changing environmental conditions. However, the extraction and analysis of organic acids can be challenging and validated methods available in this field are limited. In this study, we developed a method for the extraction and quantification of organic acids from microbial samples based on solid-phase extraction on a strong anionic exchange cartridge and gas chromatographic-mass spectrometric analysis. Full method validation was conducted to determine quality parameters of the new method. Recoveries for 12 of the 15 aromatic and aliphatic acids were between 100 and 111% and detection limits between 3 and 272 ng/mL. The ranges for the regression coefficients and process standard deviations for these compound classes were 0.9874-0.9994 and 0.04-0.69 µg/mL, respectively. Limitations were encountered when targeting aliphatic acids with hydroxy, oxo or enol ester functions. Finally, we demonstrated the applicability of the method on cell extracts of the bacterium Escherichia coli and the dinoflagellate Prorocentrum minimum. Graphical abstract.

Interactions of insulin with tragacanthic acid biopolymer: Experimental and computational study.

Alternative methods for insulin delivery instead of subcutaneous injection in diabetic patients is of great essential, and biocompatible polymers are one of the most efficient vehicles for this purpose. This research aims to investigate the capability of tragacanthic acid (TA) to bind insulin and release it under physiological conditions without alteration in the structure and conformation of insulin. Interactions between TA and insulin were studied using spectroscopic techniques and computational modeling by docking and molecular dynamics simulations. Our results demonstrate an entropy-driven spontaneous interaction between insulin and TA, where hydrogen bonds act as the main enthalpic contribution. According to our findings, the weak interaction between insulin and TA provides the basis for efficient capture and appropriate release of insulin by TA as a potential part of the insulin delivery system. In conclusion, tragacanth acid can be a proper candidate for insulin delivery.

The use of a double coaxial electrospray ionization sprayer improves the peak resolutions of anionic metabolites in capillary ion chromatography-mass spectrometry.

Recently, ion chromatography coupled with mass spectrometry has been used for the determination of anionic metabolites. However, connection with a mass spectrometer in this method is not straightforward because backpressure produced by the addition of a make-up solution often affects the peak resolutions of the target metabolites. To overcome this problem, we developed a capillary ion chromatography-mass spectrometry method utilizing a double coaxial electrospray ionization sprayer. This method was not affected by backpressure and the number of theoretical plates was about three times that of a conventional sprayer. Under optimized conditions, 44 anionic metabolites, including organic acids, sugar phosphates, nucleotides, and cofactors, were successfully separated and selectively detected with a Q Exactive mass spectrometer. The calibration curves of the tested metabolites showed excellent linearity within the range of 1-100,000 nmol/L and the correlation coefficient was greater than 0.991. The detection limits for these metabolites were between 1 and 500 nmol/L (0.4 and 200 fmol). The developed method was applied to the quantitation of anionic metabolites in cultured cancer cell samples with tumor necrosis factor (TNF)-α stimulation. This allowed for the successful determination of 105 metabolites. The levels of tricarboxylic acid cycle intermediates changed significantly after TNF-α stimulation. These results demonstrate that the developed method is a promising new tool for comprehensive analysis of anionic metabolites.

Cuban Sugar Cane Wax Acid and Policosanol Showed Similar Atheroprotective Effects with Inhibition of LDL Oxidation and Cholesteryl Ester Transfer via Enhancement of High-Density Lipoproteins Functionality.

BACKGROUND: Cuban sugarcane wax acids (SCWA) and policosanol (PCO) are mixtures of higher aliphatic acids and alcohols, respectively, purified from sugarcane wax with different chief components. Although it has been known that they have antioxidant and anti-inflammatory activities, physiological properties on molecular mechanism of SCWA have been less studied than PCO. METHODS: In this study, we compared antiatherogenic activities of SCWA and PCO via encapsulation with reconstituted high-density lipoproteins (rHDL). RESULTS: After reconstitution, SCWA-rHDL showed smaller particle size than PCO-rHDL with increase of content. PCO-rHDL or SCWA-rHDL showed distinct inhibition of glycation with similar extent in the presence of fructose. PCO-rHDL or SCWA-rHDL showed strong antioxidant activity against cupric ion-mediated oxidation of low-density lipoproteins (LDL), and inhibition of oxLDL uptake into macrophages. Although PCO-rHDL showed 1.2-fold stronger inhibition against cholesteryl ester transfer protein (CETP) activity than SCWA-rHDL, SCWA-rHDL enhanced 15% more brain cell (BV-2) growth and 23% more regeneration of tail fin in zebrafish. CONCLUSION: PCO and SCWA both enhance the beneficial functions of HDL to maximize its antioxidant, antiglycation, and antiatherosclerotic activities and the inhibition of CETP. These enhancements of HDL functionality by PCO and SCWA could exert antiaging and rejuvenation activity.

Chromatographic and thermodynamic comparison of amylose tris(3-chloro-5-methylphenylcarbamate) coated or covalently immobilized on silica in high-performance liquid chromatographic separation of the enantiomers of select chiral weak acids.

The separation of enantiomers of some chiral weak acids was studied in HPLC with chiral HPLC columns prepared by coating or covalent immobilization of the same chiral selector, namely amylose tris(3-chloro-5-methylphenylcarbamate) onto silica. After screening some representatives of arylpropionic acid derivatives, coumarins and barbiturates in hydrocarbon-alcohol type mobile phases, we studied the temperature dependence of separation parameters for ketoprofen and naproxen. Instances of reversal of the enantiomer elution order were observed function of column temperature, nature of polar modifier and its content in the mobile phase, as well as between the coated and covalently immobilized versions of the columns made with more-or-less the same chiral selector. Thermodynamic parameters such as Gibb's free energy, the standard molar entropy and the standard molar enthalpy of analyte transfer from the mobile to the stationary phase were calculated in some cases in order to explain the differences observed in the enantiomer separation ability and pattern of coated and covalently immobilized columns.

Arvensic acids A-D, novel heptasaccharide glycosidic acids as the alkaline hydrolysis products of crude resin glycosides from Convolvulus arvensis.

In this study, a resin glycoside fraction with cytotoxic activity was isolated from the alcoholic extract of C. arvensis whole plants. To describe the chemical feature of the resin glycosides, the fraction was alkaline hydrolyzed and four novel glycosidic acids, named arvensic acids A-D (1-4), were isolated. Their structures were thoroughly elucidated on the basis of spectroscopic data and chemical evidences. They all possess a same heptasacharride core, consisting of one D-fucose, two L-rhamnose and four d-glucose units. The difference among these glycosidic acids was placed on the aglycone, which is 12S-hydroxypentadecanoic acid for 1, 12S-hydroxyhexadecanoic acid for 2, 3S,12S-dihydroxypentadecanoic acid for 3, and 3S,12S- dihydroxyhexadecanoic acid for 4. These aglycones are rarely found in the structures of resin glycosides and are firstly identified in the genus Convolvulus.

Matured Hop-Derived Bitter Components in Beer Improve Hippocampus-Dependent Memory Through Activation of the Vagus Nerve.

Improving and maintaining memory function is effective in preventing cognitive decline and dementia. Previously, we demonstrated that iso-α-acids, the hop-derived bitter components in beer, prevent cognitive impairment in an Alzheimer's disease mouse model. In this report, we investigated the effects of matured hop bitter acids (MHBA) containing components of oxides derived from α- and ß-acids, and structurally similar to iso-α-acids, on cognitive function using behavioral pharmacological procedures. MHBA and the representative components of MHBA, 4'-hydroxyallohumulinone (HAH) and 4'-hydroxy-cis-alloisohumulone (HAIH) improved spatial working memory in scopolamine-induced amnesia mice. MHBA also enhanced episodic memory in the novel object recognition test (NORT). The administration of MHBA increased the amount of norepinephrine (NE) and NE release into cerebrospinal fluid (CSF) in hippocampus. The MHBA activity in improving memory function was attenuated by treatment with a ß-adrenergic receptor inhibitor. In addition, vagotomized mice did not display the memory improvement induced by MHBA. Together, our results suggest that MHBA improves memory function via stimulation of the vagus nerve and enhancement of NE release in the hippocampus. Vagus nerve activation by the intake of food materials including MHBA may be a safe and effective approach for improving cognitive function.

Separation of Benzoic and Unconjugated Acidic Components of Leonardite Humic Material Using Sequential Solid-Phase Extraction at Different pH Values as Revealed by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry and Correlation Nuclear Magnetic Resonance Spectroscopy.

Here, we report on sequential solid-phase extraction of leonardite hymatomelanic acid (CHM) on a non-ionic sorbent at four steadily lowered pH values: 7, 5, 3, and 2, yielding fractions with different acidic properties. Using nuclear magnetic resonance (NMR) spectroscopy and ultrahigh-resolution mass spectrometry, we revealed a gradual shift of dominating scaffolds in the fractions of CHM from reduced saturated to oxidized aromatic compounds. An increase on the average aromaticity of the CHM fractions was accompanied by a red shift in fluorescence spectra. These results were supported by heteronuclear single quantum coherence and heteronuclear multiple bond correlation NMR experiments. We have demonstrated that the CHM fraction isolated at pH 5 was dominated by aliphatic carboxyl carriers, while the pH 3 fraction was dominated by aromatic carboxyl acids. The developed fractionation technique will enable deeper insight on structure-property relationships and the design of the humic-based materials with tailored reactive properties.

Isolation of acid from eye drop bottles being used by patients presenting with presumed scleritis.

Purpose: The aim of the study was to report the occurrence of contamination/replacement of ophthalmic eye drops with liquids of acidic nature in patients treated for nonresponding scleritis. Methods: This was a retrospective interventional case series study. Results: Of the three patients (4 eyes) referred as necrotizing scleritis, two were found to have acid as the content in the bottle/s being used as eye drops, confirmed using biochemical tests. All four eyes had tarsal ischemia and tarsal conjunctival defect in addition to severe scleral ischemia involving the inferior bulbar area. All four eyes required tenonplasty with amniotic membrane transplant more than once for the ocular surface to heal. Two of the three patients were on systemic immunosuppressives including pulse cyclophosphamide for refractory necrotizing scleritis. Sulfuric and hydrochloric acid was isolated from the bottles of 2nd and 3rd patient using confirmatory biochemical tests. Conclusion: It is important to be aware of the possibility of contaminating or replacing contents of eye drops with harmful agents of acidic nature and should be considered in situations that resemble the clinical picture described herein.

Preparation and evaluation of surface-bonded phenylglycine zwitterionic stationary phase.

4-Hydroxy-D-phenylglycine was modified with methacrylic anhydride and then immobilized on silica through thiol-initiated surface polymerization; the prepared material was applied as stationary phase for HPLC. The obtained stationary phase was characterized by elemental analysis, infrared spectroscopy, and thermogravimetric analysis. The chromatographic performance of the packed column was evaluated in reversed-phase liquid chromatograph (RPLC) and hydrophilic interaction liquid chromatograph (HILIC) mode; this column has shown excellent selectivity to both the hydrophobic and hydrophilic solutes. The selectivity towards polycyclic aromatic hydrocarbons relative to that towards alkylbenzenes exhibited by the prepared column was higher than the corresponding selectivity exhibited by commercial C18 column, which could be explained by electronic π-π interaction between phenylglycine and electron-rich aromatic rings. On the other hand, the prepared column has also shown better selectivity for polar compounds, which was based on the multiple interaction and retention mechanisms. It was also used to separate sulfonamides and organic acid compared with a commercial C18 and HILIC column; the results show its great chromatographic performance with distinctive selectivity. All the results indicated the prepared column had potential application in a wide range.

Demonstration of acid and water recovery systems: Applicability and operational challenges in Indian metal finishing SMEs.

Diffusion dialysis, acid retardation and nanofiltration plants were acquired from Europe and demonstrated in several Indian metal finishing companies over a three year period. These companies are primarily small and medium enterprises (SMEs). Free acid recovery rate from spent pickling baths using diffusion dialysis and retardation was in the range of 78-86% and 30-70% respectively. With nanofiltration, 80% recovery rate of rinse water was obtained. The demonstrations created awareness among the metal finishing companies to reuse resources (acid/water) from the effluent streams. However, lack of efficient oil separators, reliable chemical analysis and trained personnel as well as high investment cost limit the application of these technologies. Local manufacturing, plant customization and centralized treatment are likely to encourage the uptake of such technologies in the Indian metal finishing sector.

Capillary electrophoresis in association with chemometrics approach for bitterness hop (Humulus lupulus L.) classification.

The precursor compounds related to the bitterness of beer are called α-acids. These compounds are extracted from the hop, which is an important ingredient in the brewing process. These compounds were analyzed by capillary electrophoresis. The electrophoretic method used 160 mmol/L of ammonium carbonate (pH 9) as BGE (background electrolyte), a voltage of +20 kV in a capillary with 50 µm of internal diameter and with a 62.5 cm of total length (54 cm effective). The samples were injected in hydrodynamic mode applying a pressure of 25 mbar for 5 s and the analytes were detected at 230 nm. A hydromethanolic extraction during 3 h was considered as the optimum condition for the sample preparation using MeOH/H2 O 80:20 v/v as the extract solution. From the optimized conditions the electropherograms were evaluated for their use as input for chemometric modeling. Preprocessing investigation for electrophoretic data taking into account the alignment, denoising and baseline correction, and variable selection were considered before the chemometric modeling using principal component analysis (PCA). The electrophoretic data were systematically evaluated to find the optimum conditions to modeling. A PCA analysis for all tests was carried out using different preprocessing methods and, an explained variance higher than 90% was achieved in all of them. The optimized chemometric method worked with aligned and meancentered data. From this approach, a simple and efficient method to classify hop samples with high and low α-acids content without the use of analytical standards was established from a simple electrophoretic analysis.

Experiment on the treatment of acid mine drainage with optimized biomedical stone particles by response surface methodology.

The immobilized particles were used to treat acid mine drainage (AMD) in the study, which owns the characteristics of serious pollution and high managing cost. The immobilized particles were prepared with sulfate reducing bacteria (SRB) and medical stones. In order to investigate the interactive influence of medical stones on the particle properties, the salt modification condition, content, and size of the medical stone were taken as the influential factors. At the same time, the removal rate of SO42- and Mn2+, the release of total irons (TFe) and chemical oxygen demand (COD) and pH value were taken as the response values in the experiment. On the basis of the orthogonal experimental research, a response surface model was established. The experimental analysis showed that the particles can get the best treatment effect, when using the salt-modified medical stone with the content of 15% and particle size of 200~300 mesh. At this time, the removal rates of Mn2+ and SO42- in wastewater were 83.10 and 96.22%, respectively. The release contents of TFe and COD were 2.99 mg L-1 and 1828.54 mg L-1, respectively, and the pH value was 7.05. Then, biological medical stone particles were prepared according to the optimal ratio in the response surface experiment. The adaptability of biomedical stone particles was studied at different concentrations of SO42-, Mn2+ and pH value. The results showed that the high concentration of SO42- inhibited the metabolism of SRB, while Mn2+ had a less effect. The biomedical stone particles could regulate pH value very well.

Phenylisotertronic acids from the TCM endophytic fungus Phyllosticta sp.

Four new phenylisotertronic acids (1a/1b, 2a, and 3a) were isolated from a TCM endophytic fungal strain Phyllosticta sp. J13-2-12Y obtained from the leaves of Acorus tatarinowii, along with two known ones (2b and 3b). Compounds 1-3 all existed as mixtures of enantiomers, and their corresponding optically pure enantiomers were successfully isolated by chiral HPLC. The structures of isolated compounds were determined by comprehensive spectroscopic analyses and X-ray diffraction. Their absolute configurations were determined by ECD experiments and quantum chemical calculations. In addition, the antimicrobial activities and the cytotoxicities of these three pairs of optically pure enantiomers (1a/1b, 2a/2b, and 3a/3b) had been evaluated.

Response surface methodology for the determination of the design space of enantiomeric separations on cinchona-based zwitterionic chiral stationary phases by high performance liquid chromatography.

Quality-by-Design approach for enantioselective HPLC method development surpasses Quality-by-Testing in offering the optimal separation conditions with the least number of experiments and in its ability to describe the method's Design Space visually which helps to determine enantiorecognition to a significant extent. Although some schemes exist for enantiomeric separations on Cinchona-based zwitterionic stationary phases, the exact design space and the weights by which each of the chromatographic parameters influences the separation have not yet been statistically studied. In the current work, a screening design followed by a Response Surface Methodology optimization design were adopted for enantioseparation optimization of 3 model drugs namely the acidic Fmoc leucine, the amphoteric tryptophan and the basic salbutamol. The screening design proved that the acid/base additives are of utmost importance for the 3 chiral drugs, and that among 3 different pairs of acids and bases, acetic acid and diethylamine is the couple able to provide acceptable resolution at variable conditions. Visualization of the response surface of the retention factor, separation factor and resolution helped describe accurately the magnitude by which each chromatographic factor (% MeOH, concentration and ratio of acid base modifiers) affects the separation while interacting with other parameters. The global optima compromising highest enantioresolution with the least run time for the 3 chiral model drugs varied extremely, where it was best to set low % methanol with equal ratio of acid-base modifiers for the acidic drug, very high % methanol and 10-fold higher concentration of the acid for the amphoteric drug while 20 folds of the base modifier with moderate %methanol were needed for the basic drug. Considering the selected drugs as models for many series of structurally related compounds, the design space defined and the optimum conditions computed are the key for method development on cinchona-based chiral stationary phases.