Liquid Chromatography Diode Array Detection Mass Research Articles

Towards an index of damage of olive paste during virgin olive oil extraction: The role of β-OH-acteoside diastereomers for risk assessment of the fusty/muddy sediment sensory defect

The aim of this study was to propose an index/marker of thermal/oxidative damage of olive paste during virgin olive oil extraction to assess the risk of sensory defects in the obtained oil. Several olive oil extraction trials were carried out over two production years applying different time, temperature, and oxygen exposure conditions during malaxation. The phenolic composition of the olive pastes was analyzed by High Performance Liquid Chromatography-Diode Array Detector-Mass Spectrometry (HPLC-DAD-MS). The chemical (legal quality indexes, phenolic compounds by HPLC-DAD-MS, volatile compounds by Head Space-Solid Phase Microextraction-Gas Chromatography-Mass Spectrometry (HS-SPME-GC-MS)) and sensory characteristics of the obtained oils were analyzed. The content of the β-OH-acteoside diastereomers in olive paste increased linearly with malaxation time as a function of malaxation temperature and oxidative conditions, following a pseudo-zero order kinetic. At the same time, two volatile compounds linked to sensory defects (2- and 3-methylbutanal) increased in the obtained oils when increasing malaxation time and temperature, and a relationship between them and the fusty/muddy sediment defect in olive oil samples was observed. A further direct relationship between the β-OH-acteoside content in the olive pastes and the 2+3-methylbutanal content in the oils was observed. Therefore, the β-OH-acteoside diastereomers content in olive pastes has been suggested as an index/marker to indicate the risk of sensory defects in virgin olive oil as a function of the thermal and oxidative conditions during malaxation. A threshold value of 100 mg/kg of the content of β-OH-acteoside diastereomers was proposed, above which the fusty/muddy sediment defect is perceptible.

Identification of aloe and other dyes by means of SERS and HPLC-DAD-MS in the embroidery of a 15th century English folded almanac

The folded almanac Western MS.8932 (Wellcome Collection, London), produced in England between 1387 and 1405, is a calendar with astrological tables and diagrams used by medical practitioners to harness astrological information relating to health. Apart from the great interest for its use in medicine, this object has a unique feature in its exquisite, unique embroidered binding that indicates a prestigious artefact. Considering the scarceness of information on such items, a diagnostic study on the materials used for the outstanding embroidery of MS.8932 is a unique occasion of having information on the methods of production of medieval embroideries. A diagnostic campaign has been carried out to identify the dyes used for this task and the colourants used for the illuminations, and to evaluate them with reference to their commercial value. Preliminary information on the dyes was yielded by means of UV–visible diffuse reflectance spectrophotometry with fibre optics (FORS) and spectrofluorimetry with fibre optics (FOMF). Then a final identification was yielded by micro-invasive analysis involving Surface Enhanced Raman Spectroscopy (SERS) and High Performance Liquid Chromatography-Diode Array Detector-Mass Spectrometry (HPLC-DAD-MS) [1]. Analysis on the micro samples (5–10 mm of very thin threads) taken from the embroidery revealed the presence of orchil for pink hues, indigo/madder double dyeing for purple hues, red safflower for salmon pink hues and indigo/weld double dyeing for green hues. Particularly remarkable was the additional identification of aloe in the purple threads, possibly used as mordant and/or as antibacterial agent, which evidence is the first ever recorded in such materials. This information is very important to understand the significance of the artefact and its historical-artistic location, and will allow to make comparison with similar artworks.

Does Fermentation Really Increase the Phenolic Content in Cereals? A Study on Millet.

Millet is underutilized in Europe, despite its advantages compared to other common cereals. In Asia and Africa, millet is mainly eaten in fermented form; its consumption has beneficial properties on human health. Three millet batches were compared in terms of free and bound phenols by High Performance Liquid Chromatography-Diode Array Detector-Mass Spectrometry (HPLC-DAD-MS). The richest one in terms of bound phenols was selected for testing via a basic (0.1 M NaOH) and an acidic (1.2 M H2SO4) hydrolysis, in which 149.3 and 193.6 mg/100 g of phenols were recovered, respectively. The ability of fermentation, with yeast and Lactobacilli, to increase the content of phenolic compounds was verified. Five withdrawalswere performed to verify the influence of fermentation time on the total phenolic content. The greatest phenolic content was observed after 72 h. Fermentation increased the cinnamic acids and flavonoids contents by approximately 30%. Vitexin and vitexin 2″-O-rhamnoside contents were significantly higher in the fermented millet; these compounds partially inhibit the protein tyrosine phosphatase enzyme, which is overexpressed in type-2 diabetes. A molecular dynamic simulation showed the two flavonoids to be allosteric inhibitors. The phenolic extract from fermented millet demonstrated a higher level of antioxidant protection on human erythrocytes by ex vivo cellular antioxidant activity in red blood cells. In this context, functional foods based on fermented millet could represent a new trend in European markets.

Andean Prumnopitys Andina (Podocarpacae) Fruit Extracts: Characterization of Secondary Metabolites and Potential Cytoprotective Effect.

The fruits from the Chilean Podocarpaceae Prumnopitys andina have been consumed since pre-Hispanic times. Little is known about the composition and biological properties of this fruit. The aim of this work was to identify the secondary metabolites of the edible part of P. andina fruits and to assess their antioxidant activity by means of chemical and cell-based assays. Methanol extracts from P. andina fruits were fractionated on a XAD7 resin and the main compounds were isolated by chromatographic means. Antioxidant activity was determined by means of 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), ferric reducing power (FRAP), trolox equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assays. The cytoprotective activity of the extract against oxidative and dicarbonyl stress was evaluated in human gastric epithelial cells (AGS). The total intracellular antioxidant activity (TAA) of the extract was determined in AGS cells. The inhibition of meat lipoperoxidation was evaluated under simulated gastric digestion conditions. Rutin, caffeic acid β-glucoside and 20-hydroxyecdysone were identified as major components of the fruit extract. Additional compounds were identified by high-performance liquid chromatography diode-array detector mass spectrometry (HPLC-DAD-MSn) and/or co-injection with standards. Extracts showed dose-dependent cytoprotective effects against oxidative and dicarbonyl-induced damage in AGS cells. The TAA increased with the pre-incubation of AGS cells with the extract. This is the first report on the composition and biological activity of this Andean fruit.

Determination of Four Bitter Compounds in Caramel Colors and Beverages Using Modified QuEChERS Coupled with Liquid Chromatography-Diode Array Detector-Mass Spectrometry

The 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one (DDMP), 5-hydroxymethylfurfural (HMF), furfural, and 5-methylfurfural (MF) are the components of caramel colors with intense bitterness. In this study, a practical and simple method for the simultaneous analysis of these four bitter compounds in caramel colors and beverages was proposed using high-performance liquid chromatography–diode array detector–atmospheric pressure chemical ionization–mass spectrometry (HPLC-DAD-APCI-MS). For the sample preparation, the extract efficiencies from different extract solvents and quick, easy, cheap, effective, rugged, and safe (QuEChERS) salt pockets were compared. The correlation coefficients (R2) of all analytical curves were ≥ 0.9914. The satisfactory recoveries ranged between 70.1% and 101.5%, with relative standard deviations ranged from 1.7% to 9.1%. The matrix effect was evaluated, and HMF, furfural, and MF showed signal enrichment. This method was successfully applied to the analysis of bitter compounds content in several caramel colors and beverages from a local market in China. Four compounds were found in eight samples, with concentration ranging from 0.5 to 1058.1 mg kg−1.

Extraction Optimization of Flavonoids from Hypericum formosanum and Matrix Metalloproteinase-1 Inhibitory Activity.

Hypericum formosanum is a valuable herb in Taiwan. In this study, response surface methodology was employed to optimize the ultrasound-assisted extraction of flavonoids from Hypericum formosanum. A central composite design with three variables (ethanol concentration, extraction time, and extraction temperature) was applied. Experimental results were fitted to the second order polynomial model and one-way analysis of variance was used to determine the goodness of fit of the model and the optimal conditions for responses. The optimal conditions for the maximum extraction yield of total flavonoid content (101.1 mg/g) using ultrasound-assisted extraction were ethanol concentration, 73.5%; extraction time, 38.3 min; and extraction temperature, 62.5 °C. The predicted result was consistent with the experimental result obtained under optimal extraction conditions. Hyperoside, astilbin, quercitrin, and quercetin from Hypericum formosanum extract (HFE) were identified by Ultra performance liquid chromatography-diode array detector-mass (UPLC-DAD-MS). HFE significantly reduced matrix metalloproteinase-1 protein expression in human skin keratinocyte cells, induced by advanced glycation end products.

In vitro Antioxidant and Enzymatic Approaches to Evaluate Neuroprotector Potential of Blechnum Extracts without Cytotoxicity to Human Stem Cells.

Background:Investigation of selected plant extracts on multi-targets related to neurodegeneration, such as monoamine oxidases (MAO), cholinesterase enzymes, and antioxidant activities (AOA) is a useful tool for identification of new scaffolds.Objective:This work investigated biological effects of three Blechnum methanol extracts from Brazil and chemical profile of the most active sample.Materials and Methods:AOA included scavenging of hydroxyl and nitric oxide radicals, also lipid peroxidation inhibition. Enzymatic modulation of Blechnum binervatum, Blechnum brasiliense, and Blechnum occidentale extracts on MAO and cholinesterases was conducted. Moreover, total phenol content was performed with all samples, and high-performance liquid chromatography-diode array detection mass spectrometry HPLC-DAD-MS analysis was carried out with B. brasiliense. Possible toxic effects were evaluated on Wistar rats polymorphonuclear cells (PMN) and human stem cells.Results:B. brasiliense extract presented the highest phenolic amount (9.25 g gallic acid equivalent/100 g extract) and lowest IC50 values (112.3 ± 2.61 and 176.1 ± 1.19 μg/mL) against hydroxyl radicals and on lipid peroxidation, respectively, showing strong AO effects. On nitric oxide assay and cholinesterase inhibition, all extracts were considered inactive. MAO-A selective action was evidenced, being B. brasiliense powerful against this enzyme (IC50: 72.7 μg/mL), followed by B. occidentale and B. binervatum (IC50: 130.85 and 165.2 μg/mL). No cytotoxic effects were observed on PMN and human stem cells treated with Blechnum extracts. HPLC-DAD-MS analysis of B. brasiliense allowed the identification of chlorogenic and rosmarinic acids.Conclusion:Our results especially highlight B. brasiliense, with pronounced phenols content and strong effects on selected targets related to neurodegeneration, being characterized as a natural safe source of bioactive hydroxycinnamic acids.SUMMARY Blechnum crude extracts showed high phenolic amounts and valuable IC50 values on targets related with neurodegenerative disordersBlechnum brasiliense was the most active sample, with strong radical scavenging and lipid peroxidation inhibition, also with monoamine oxidases: A selective modulationNo cytotoxic effects were observed on polymorphonuclear cells rat cells and human stem cells treated with Blechnum extractsHigh-performance liquid chromatography-diode array detection-mass spectrometry analysis of Blechnum brasiliense allowed the identification of hydroxycinnamic derivatives: Chlorogenic and rosmarinic acids. Abbreviations used: IC50: half maximal inhibitory concentration; MAO: monoamine oxidase; MAO-A: monoamine oxidase isoform A; MAO-B: monoamine oxidase isoform B; HO•: hydroxyl radical.

Qualitative and quantitative analysis of an alkaloid fraction from Piper longum L. using ultra-high performance liquid chromatography-diode array detector–electrospray ionization mass spectrometry

In a previous research, an alkaloid fraction and 18 alkaloid compounds were prepared from Piper longum L. by series of purification process. In this paper, a qualitative and quantitative analysis method using ultra-high performance liquid chromatography-diode array detector–mass spectrometry (UHPLC-DAD–MS) was developed to evaluate the alkaloid fraction. Qualitative analysis of the alkaloid fraction was firstly completed by UHPLC-DAD method and 18 amide alkaloid compounds were identified. A further qualitative analysis of the alkaloid fraction was accomplished by UHPLC–MS/MS method. Another 25 amide alkaloids were identified according to their characteristic ions and neutral losses. At last, a quantitative method for the alkaloid fraction was established using four marker compounds including piperine, pipernonatine, guineensine and N-isobutyl-2E,4E-octadecadienamide. After the validation of this method, the contents of above four marker compounds in the alkaloid fraction were 57.5mg/g, 65.6mg/g, 17.7mg/g and 23.9mg/g, respectively. Moreover, the relative response factors of other three compounds to piperine were calculated. A comparative study between external standard quantification and relative response factor quantification proved no remarkable difference. UHPLC-DAD–MS method was demonstrated to be a powerful tool for the characterization of the alkaloid fraction from P. longum L. and the result proved that the quality of alkaloid fraction was efficiently improved after appropriate purification.

Metabolite identification of a new tyrosine kinase inhibitor, HM781‐36B, and a pharmacokinetic study by liquid chromatography/tandem mass spectrometry

HM781-36B (1-[4-[4-(3,4-dichloro-2-fluorophenylamino)-7-methoxyquinazolin-6-yloxy]-piperidin-1-yl]prop-2-en-1-one hydrochloride) is a new anticancer drug to treat advanced solid tumors in clinical trial. In order to understand the behavior of HM781-36B in vitro and in vivo we validated an analytical method for HM781-36B and its major metabolites in plasma. In vivo and in vitro metabolism of HM781-36B was studied in dog plasma, urine and feces as well as using human and dog liver microsomes with extraction by ethyl acetate or methyl tert-butyl ether, respectively, and successfully separated by high-performance liquid chromatography diode-array detection mass spectrometry (HPLC-DAD/MS). Ten metabolites were identified by LC/ESI-ion trap mass spectrometry (MS, MS(2) , MS(3) and MRM) and LC/Q-TOF-MS/MS for exact mass measurement. For accurate characterization of the major metabolites, authentic standards (M1, M2, M4, and M10) were synthesized. Ten metabolites of HM781-36B in an in vitro mixture were separated and identified by LC/ESI-MS(n) . The MS/MS spectral patterns of the parent drug and metabolites exhibited two characteristic ions (A- and B-type ions) attributed to the cleavage of the ether bond between the piperidine ring and the quinazoline ring, providing important information on the site of chemical conversion during the metabolism. Six hydroxylated derivatives including dehalogenation and demethylation, two N-oxide forms, a demethylated form and de-acryloylpiperideine metabolites were observed. The LC/ESI-ion trap MS(n) technique was effective in obtaining structural information and yielded diagnostic ions for the identification of diverse metabolites. The multiple metabolic pathways of HM781-36B were suggested in in vitro and in vivo samples and the dihydroxylation (M1) and demethylation (M2) appeared to be the major metabolites.

Qualitative and Quantitative Analysis of Swertia Herbs by High Performance Liquid Chromatography-Diode Array Detector-Mass Spectrometry (HPLC-DAD-MS)

We evaluated the composition of Swertia herbs using high performance liquid chromatography-diode array detector-mass spectrometry (HPLC-DAD-MS). Eleven peaks of 6 species were unequivocally identified by comparing their retention times, UV spectra, on-line electrospray ionization mass (ESI-MS) spectra, and collision-induced dissociation mass spectrometry/mass spectrometry (CID-MS/MS) data with those of authentic compounds. We adopted wavelengths of 254 nm, 340 nm and 230 nm to simultaneously determine these 11 compounds. By comparing the overall DAD and total ion current (TIC) profiles of various samples, the 6 species were differentiated in terms of the occurrence and/or relative concentrations of the eleven compounds. Our novel validated HPLC-DAD-MS method not only facilitates quality control and identification of Swertia herbs, but is also applicable to systematic investigations of the distribution of secoiridoids, flavonoids, and xanthones in the genus Swertia.

Analysis of environmental samples by application of multivariate curve resolution on fused high-performance liquid chromatography–diode array detection mass spectrometry data

Multivariate curve resolution-alternating least squares (MCR-ALS) is applied to solve coelution problems in liquid chromatograpy with diode array detection (DAD) and mass spectrometry (MS). Fusion of DAD and MS detector signals improved results versus those obtained using only one of the two detector signals. Application of wavelet transform to MS data before its fusion with DAD data, further help to facilitate the resolution and quantitation of the coeluted compounds under study, besides a decrease of time of analysis. Mixtures of biocide compounds in standard mixtures and in environmental samples (sediment and wastewater samples) were analyzed with estimated quantitation errors below 12%.