Array Detection Mass Spectrometry Research Articles

Valorisation of Winery By-Products: Revealing the Polyphenolic Profile of Grape Stems and Their Inhibitory Effects on Skin Aging-Enzymes for Cosmetic and Pharmaceutical Applications.

Grape (Vitis vinifera L.) stems, a by-product of winemaking, possess significant potential value due to their rich polyphenolic composition, which allows their exploitation for cosmetic and pharmaceutical applications. This presents a promising opportunity for valorisation aimed at developing innovative products with potential health-promoting effects. In this study, the polyphenolic profile of extracts from grape stems of seven white grape varieties was determined using spectrophotometric and chromatographic methods, specifically high-performance liquid chromatography coupled with a photodiode array detector and electrospray ionization multi-stage mass spectrometry (HPLC-PDA-ESI-MSn), as well as on their ferric-reducing antioxidant power (FRAP) and radical scavenging capacity, using 2,2-diphenyl-1-picrylhydrazyl (DPPH●) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS●+) radicals. This study also evaluated the anti-aging activity and skin depigmenting activity of these extracts. These findings revealed a diverse polyphenolic profile, encompassing proanthocyanidins and catechin derivatives (PCDs), phenolic acids, and flavonols. Among the varieties studied, 'Códega do Larinho' exhibited the highest concentrations of six distinct polyphenols and the highest total phenolic content. It also demonstrated the highest results for antioxidant capacity and elastase and tyrosinase inhibition. Pearson's correlation analysis showed a significant positive correlation between certain PCDs with both FRAP and DPPH assays, as well as between the identified flavonols and anti-elastase activity. These results underscore the potential health benefits of grape stem extracts and emphasize the importance of their polyphenolic composition in enhancing antioxidant and anti-aging properties, thus supporting their application in different industries.

“Welcome to the jungle”: TLC-SERS to wade through real complex mixtures of synthetic dyes

The finding of numerous synthetic strategies between the 19th and 20th century fostered the production of an extensive spectrum of organic dyes for artistic and industrial applications. Shortly after, challenges emerged related to the rapid deterioration of these products. Nowadays, the characterization of coloured pieces of art produced in the first half of the XXth century is further complicated by the absence of reference materials with an unambiguous composition, which are frequently a mixture between the starting dyes and their degradations products. Here we demonstrate as Surface Enhanced Raman Spectroscopy coupled with the simplest separation technique, i.e. Thin Layer Chromatography, is able beyond to recognize several synthetic dyes also to identify their isomers and degradation products. The occasion was the study of a catalogue of dyed textile swatches, produced in 1951 by the well-known Italian industry ACNA (Azienda Coloranti Nazionali e Affini). An apparently easy task turned out to be, indeed, not at all obvious as we found out that the actual composition of the reference samples seldomly corresponded to the declared one. TLC-SERS investigation of historical textile swatches was complemented by the integration of liquid chromatography coupled with a diode array detector and high-resolution mass spectrometry (HPLC-DAD-HRMS). This approach allowed us to validate the findings of TLC-SERS and to distinctively characterise synthetic by-products and degradation markers resulting from the deterioration of the dyes.

Gastrointestinal digestion of olive leaf waste from olive mill production chain in a simulator of the gastrointestinal tract.

Currently, olive leaves are considered waste, although their high phenolic content makes them a source of antioxidants that could be used directly. The aim of this work was to study the behavior of phenolic compounds contained in olive leaf from the olive mill production chain during their gastrointestinal digestion. Phenolic compounds in the gastric digestion analyzed by high-performance liquid chromatography-diode array detection-mass spectrometry increased by 58% to 314.5% compared to the meal, while in the intestinal stage they ranged from 1.87 to 9.04 times higher. An increase of between 187% and 903% in bioaccessibility of phenolic compounds in the intestinal phase was observed, except for verbascoside. Furthermore, such compounds were fully bioavailable, except for apigenin-7-O-glucoside, which showed a bioavailability of 56%. The current study showed that the concentration of oleuropein, apigegin-7-O-glucoside, hydroxytyrosol hexoside and hydroxytyrosol contained in olive leaf continued to be extracted during the gastrointestinal digestion process. Furthermore, the results obtained with respect to their bioaccessibility and bioavailability suggest a good disposition to pass into the bloodstream where they could exert beneficial effects. Therefore, these results are promising for olive leaf becoming a consumable by-product that could be directly ingested through a simple infusion. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Ultrasound-based strategies for the recovery of microalgal carotenoids: Insights from green extraction methods to UV/MS-based identification

Carotenoids, versatile natural pigments with numerous health benefits, face environmental concerns associated with conventional petrochemical-based extraction methods and limitations of their synthetic equivalents. In this context, this study aims to introduce eco-friendly approaches using ultrasound-based strategies (probe and bath) for the extraction of carotenoids from microalgae, initially focusing on Microchloropsis gaditana and subsequently evaluating the versatility of the method by applying it to other microalgae species of interest (Tisochrysis lutea, Porphyridium cruentum, and Phaeodactylum tricornutum) and defatted microalgal residues. Among the approaches evaluated, the 5-min ultrasonic probe system with ethanol showed comparable carotenoid recovery efficiency to the reference method (agitation, 24 h, acetone) (9.4 ± 2.5 and 9.6 ± 3.2 mg g−1 carotenoids per dry biomass, for the green and the reference method, respectively). Moreover, the method's sustainability was demonstrated using the AGREEprep™ software (scored 0.62 out of 1), compared to the traditional method (0.22 out of 1). The developed method yielded high carotenoid contents across species with diverse cell wall compositions (3.1 ± 0.2, 2.1 ± 0.3, and 4.1 ± 0.1 mg g−1 carotenoid per dry biomass for T. lutea, P. cruentum, and P. tricornutum, respectively). Moreover, the application of the method to defatted biomass showed potential for microalgal valorization with carotenoid recovery rates of 41 %, 60 %, 61 %, and 100 % for M.gaditana, P. tricornutum, T. lutea, and P. cruentum, compared to the original biomass, respectively. Furthermore, by using high-performance liquid chromatography with a diode array detector (HPLC-DAD) and high-resolution mass spectrometry (HRMS), we reported the carotenoid and chlorophyll profiles of the different microalgae and evaluated the impact of the eco-friendly methods. The carotenoid and chlorophyll profiles varied depending on the species, biomass, and method used. In summary, this study advances a green extraction method with improved environmental sustainability and shorter extraction time, underscoring the potential of this approach as a valuable alternative for the extraction of microalgal pigments.

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.

Chemical characterization, multivariate analysis and comparison of biological activities of different parts of Fraxinus mandshurica.

Fraxinus mandshurica (Oleaceae) is used as a traditional medicinal plant for the treatment of red eyes, menstrual disorders, excessive leucorrhea, chronic bronchitis and psoriasis. To perform chemical characterization of the secondary metabolites of F. mandshurica roots, bark, stems and leaves, 32 samples were collected from eight provinces in this study. A total of 64 chemical components were detected from four different parts of F. mandshurica by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. Meanwhile, a total of nine secoiridoids were obtained by natural product chemical extraction, isolation and identification methods. Quantitative analysis by high-performance liquid chromatography-diode array detection-mass spectrometry showed the highest total content of secoiridoids in the bark, which is also consistent with the traditional medicinal parts. The results of methodological validation showed that the correlation coefficient (R2) values were all >0.9993, indicating a good linear range of the standard curve, while the relative standard deviations of precision, reproducibility and stability were <3%, and the spiked recoveries ranged from 98.22 to 102.27%, indicating that the experimental method was reliable and stable. In addition, fingerprinting and a heatmap were established to demonstrate the content trends of F. mandshurica more visually from different origins. Multivariate analysis, including principal component analysis and partial least squares discriminant analysis, was performed to determine the chemical characteristics of different parts of F. mandshurica, and six characteristic secoiridoids that could be used to distinguish different origins were screened. Finally, the inhibition of tyrosinase, α-glucosidase, acetylcholinesterase and pancreatic lipase activities by the nine characteristic compounds and extracts from different parts were investigated, and the results showed that they all exhibited different degrees of enzyme activity inhibition and thus have potential applications in whitening and blemish removal, hypoglycemia, anti-Alzheimer's disease and anti-obesity as a new source of natural enzyme activity inhibitors. This study establishes an identification and evaluation method applicable to phytochemistry of different origins, which is a guideline for quality control, origin evaluation and clinical application of traditional medicinal plants. This is also an unprecedented study on the identification of the chemical composition of different parts of F. mandshurica, characteristic compounds and the inhibition of enzyme activity of extracts from different parts.

Simultaneous Quantification of Nine Target Compounds in Traditional Korean Medicine, Bopyeo-Tang, Using High-Performance Liquid Chromatography-Photodiode Array Detector and Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry.

Bopyeo-tang (BPT) is composed of six medicinal herbs (Morus alba L., Rehmannia glutinosa (Gaertn.) DC., Panax ginseng C.A.Mey., Aster tataricus L.f., Astragalus propinquus Schischkin, and Schisandra chinensis (Turcz.) Baill.) and has been used for the treatment of lung diseases. This study focused on establishing an analytical method that can simultaneously quantify nine target compounds (i.e., hydroxymethylfurfural, mulberroside A, chlorogenic acid, calycosin-7-O-glucoside, 3,5-dicaffeoylquinic acid, quercetin, kaempferol, schizandrin, and gomisin A) from a BPT sample using high-performance liquid chromatography with a photodiode array detector (HPLC-PDA) and ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS). The separation of compounds in both analyses was performed on a C18 reversed-phase column using the gradient elution of water-acetonitrile as the mobile phase. In particular, the multiple reaction monitoring mode was applied for quick and accurate detection in UPLC-MS/MS analysis. As a result of analyzing the two methods, HPLC-PDA and UPLC-MS/MS, the coefficient of determination of the regression equation for each compound was ≥0.9952, and recovery was 85.99-106.40% (relative standard deviation (RSD) < 9.58%). Precision testing of the nine compounds was verified (RSD < 10.0%). The application of these analytical assays under optimized conditions for quantitative analysis of the BPT sample gave 0.01-4.70 mg/g. Therefore, these two assays could be used successfully to gather basic data for clinical research and the quality control of BPT.

Holotrichones A and B, potent anti-leukemic lindenane-type sesquiterpene trimers with unprecedented complex carbon skeletons from a rare Chloranthus species

Two lindenane-type sesquiterpene (LDS) trimers with unprecedented carbon skeletons, holotrichones A (1) and B (2), were obtained from the whole plant of Chloranthus holostegius var. trichoneurus by a ultra performance liquid chromatography-photodiode array detector-mass spectrometry (UPLC-PDA-MS)-guided isolation strategy. Compound 1 represents the first LDS trimer incorporating a unique 3/5/6/6-fused framework, in which a lindenane-type monomer and the 2-methylbutyryl substituent of an LDS dimer is bridged by a six-membered ring system. Compound 2 is the first hetero-trimer fused by an LDS dimer with a p-benzoquinone-meroterpenoid, featuring an unusual 3/5/6/6/3/5/6/6/6 nonacyclic system fused by the sesquiterpenoid unit and a 2-geranyl-6-methyl-2,5-cyclohexadien-1,4-dione moiety. In compound 2, the dimeric LDS moiety is equipped with a rare oxaspiro[4.5]decane system. Their structures, including absolute configurations, were established by spectroscopic methods, GIAO NMR calculations and DP4+ probability analyses, electronic circular dichroism (ECD) calculations, and single-crystal X-ray diffraction analysis. The plausible biogenetic pathway speculation indicated that hetero- and homo-Diels-Alder additions may dominate the formation of these highly fused polycyclic frameworks. Both compounds 1 and 2 induced the human acute myeloid leukemia MV-4–11 cell death via apoptosis induction, which deserves further investigation on this new chemical class of LDS oligomers for their anti-leukemic potential.

New insights into the dyes of Central Asian ikat textiles

Central Asian ikat textiles are characterised by their bold and large abstract patterns, made up of vibrant colours with a characteristic “blurred edge” effect, which makes them some of the most recognisable fabrics worldwide. Eleven ikats from the collection of the National Museum of Asian Art, Smithsonian Institution (Washington, DC, USA) and six ikats from the collection of the British Museum (London, UK) were investigated by high pressure liquid chromatography coupled to diode array detector and high-resolution tandem mass spectrometry (HPLC-DAD-HRMS/MS) to identify the dyes and expand on recent research focused on the integration of synthetic dyes in ikat dyeing during the second half of the 19th century. The results confirmed that the palette of natural dyes includes cochineal (probably mostly from a local species native to central Asia) for red, larkspur (Delphinium semibarbatum) for yellow, and indigo for blue as the main dye sources. In some of the textiles, madder (Rubia tinctorum) for red and pagoda tree (Sophora japonica) for yellow were also detected as minor components mixed with cochineal and larkspur, respectively. Synthetic dyes, such as scarlet reds (C.I. 16150 and 16155), rose bengale (C.I. 45435), eosin (C.I. 45380), erythrosine (C.I. 45430), fuchsin (C.I. 42510/42520), methyl violet (C.I. 42535/42555), malachite green (C.I. 42000) and brilliant green (C.I. 42040) were detected in eight textiles. In three textiles, natural dyes were identified in the ikat-dyed silk warp threads and synthetic dyes in the red cotton weft threads, highlighting interesting mixtures of materials and textile-making practices. Natural yellow dyes were used in all textiles, even those for which all the other colours are synthetic. These results provide significant new information about the changes in ikat production that occurred during the 19th century in Central Asia.

On the reliability of historic books as sources of reference samples of early synthetic dyes – The case of “The Coal Tar Colours of the Farbwerke vorm. Meister, Lucius & Brüning, Höchst on the Main, Germany – A General Part” (1896)

The swatches present in the popular book “The Coal Tar Colours of the Farbwerke vorm. Meister Lucius & Brüning, Höchst on the Main, Germany – A General Part”, published in 1896, were analysed by high performance liquid chromatography coupled to diode array detector and high-resolution tandem mass spectrometry (HPLC-DAD-MS/MS) with the aim to identify the synthetic dyes and reveal the correspondences and discrepancies between the information reported in the labels and the actual chemical composition of the samples.The research resulted in guidelines to be followed to connect commercial dye names with Colour Index (C.I.) constitution numbers and produced a molecular database of more than 250 molecules, whose high-resolution mass spectrometric details are reported here for the first time in some cases. Out of 120 samples analysed, ca. 40% did not exactly correspond to the expected dye(s) according to the labels, thus raising awareness of the challenges related to the reliability of historic sources of reference materials and the need to study the composition of these historic samples before using them for chemical research purposes. Furthermore, molecular databases of synthetic dyes are fundamental for the correct identification of these materials in historic textiles, objects and works of art. Thus, the results of this research are extremely useful to all researchers studying dyed objects produced from the second half of the 19th century onwards.

Quantitative determination of specialised metabolites in different parts of Juglans regia Linn. and Carya illinoinensis (Wangenh.) K. Koch by using UHPLC-DAD-QTOF-MS/MS.

Juglans regia Linn. and Carya illinoinensis (Wangenh.) K. Koch are nut-producing plant species of the Juglandaceae family. Bioactive compounds like naphthoquinones, tetralones, and diarylheptanoids are dominant in these species. The study aimed to develop and validate a fast and sensitive analytical method by ultrahigh-performance liquid chromatography diode array detection mass spectrometry (UHPLC-DAD-MS) for quantification and identification of bioactive compounds in fruit pericarps and leaves of J.regia and C.illinoinensis collected from two different states of north India. The dried pericarps of J.regia and C.illinoinensis (500 mg) were extracted with ethyl acetate-methanol (50:50 v/v, 20 mL, 50°C, 30 min) by ultrasonication and analysed by ultrahigh-performance liquid chromatography diode array detection quadrupole time-of-flight tandem mass spectrometry (UHPLC-DAD-QTOF-MS/MS) for qualitative and quantitative examination of phytoconstituents. The method was validated according to International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human use (ICH) guidelines for linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ). Here, we report the quantification of dihydrophaseic acid (1), 4,5-dihydroxytetralone (2), 4,8-dihydroxytetralone (3), 5,8-dihydroxy-3-methoxytetralone (4), and juglanin A (5) in the pericarps and leaves of J.regia and C.illinoinensis. Furthermore, using the hyphenated analytical method, a total of 62 compounds were tentatively characterised in different samples. The principal component analysis (PCA) showed diversity between the analysed sample's composition. Also, the study evaluated the variation of bioactive compounds among different parts of J.regia and C.illinoinensis collected from different regions of northern India by UHPLC-DAD-QTOF-MS/MS. The developed method is simple, rapid, and selective for the identification and characterisation of bioactive molecules.

Changes in the production materials of Burmese textiles in the nineteenth century—dyes, mordants and fibres of Karen garments from the British Museum’s collection

The materials of six Karen textiles from the British Museum’s collection were investigated with the main aim of studying the production changes over the course of the nineteenth century and how these changes related to local and colonial trade networks. The textiles span chronologically from the 1830s to the early 1900s according to their attribution dates, and include traditional garments such as tunics and skirts, in addition to representing a broad colour palette and different dyeing and weaving techniques. The investigation was conducted non-invasively by using digital microscopy, broadband multispectral imaging (MSI) and fibre optic reflectance spectroscopy (FORS). The results guided a sampling campaign during which samples were taken and investigated by optical microscopy (OM), scanning electron microscopy energy dispersive X-ray spectrometry (SEM–EDX) and high-performance liquid chromatography coupled to diode array detector and tandem mass spectrometry (HPLC–DAD-MS/MS). Local natural dyes, such as red and yellow from Morinda citrifolia, lac dye (Kerria lacca), gamboge (Garcinia sp.), turmeric (Curcuma longa), indigo and tannins were found in the older textiles, supporting a possible early production, whereas mixtures of natural and synthetic dyes, including synthetic alizarin (CI 58000), rhodamine B (CI 45170), orange I (CI 14600), diamond green B (CI 42000), diamond green G (CI 42040), auramine O (CI 41000), naphthol yellow S (CI 10316), chrysamine G (CI 22250), crystal violet (CI 42555) and Pigment Red 53:1 (CI 15585:1) were identified in the later textiles. Observations on mordants revealed the use of aluminium, tin and chrome, whereas fibre characterisation highlighted the use not only of cotton (Gossypium sp.) but also of felted wool imported from Europe and Chinese silk dyed with the Chinese cork tree (Phellodendron chinense) yellow dye. The results draw an interesting picture of the introduction of new dyeing materials and techniques in Myanmar over the nineteenth century, and how these impacted the production of traditional Karen textiles.

Identification and quantification of cinnamic acid derivatives in Cichorium intybus seed and its extract by High- Performance Liquid Chromatography with Diode-Array Detector (HPLC-DAD) and Electrospray Ionization Mass Spectrophotometry (LC-MS/MS)

A sensitive method coupling high-performance liquid chromatography (HPLC) with diode –array detector (DAD) and electrospray ionization mass spectrometry (MS) was optimized for separation, identification, and quantification of cinnamic acid derivatives in Cichorium intybus seed and its extract. Cinnamic acid derivatives such as chlorogenic acid, caffeic acid and Chicoric acid were quantified using respective standards. Apart from 4-o-Caffeoylqunic acid, other cinnamic acid derivative such as 3-o-caffeoylquinic acid was also identified and quantified by UV and MS/MS spectra and calculated as total caffeoylquinic acids using 4-o-caffeoylqunic acid as standard in the seed and its extract. Other cinnamic acid derivatives such as 1,3-dicaffeoylqunic acid, 1,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 3, 4-dicaffeoylquinic acid, dicaffeoylquinic acid-1 and dicaffeoylquinic acid-2 (two unknown) were identified and quantified by UV and MS spectra and calculated as total dicaffeoylquinic acids using chlorogenic acid standard in the seed and its extract. The total cinnamic acids were quantified by calculating the sum of chlorogenic acid, caffeic acid, chicoric acid, total caffeoylquinic acids(4-o-caffeoylquinic acid and 3-o-caffeoylqunic acid) and total dicaffeoyl-quinic acids(1,3-dicaffeoylqunic acid, 1,4-dicaffeoylquinic acid, 3,5-dicaffeoyl-quinic acid, 3, 4-dicaffeoylquinic acid, dicaffeoylquinic acid-1 and dicaffeoyl-quinic acid-2). The Phytochemical screening of C. intybus seed and its extract revealed that this plant is a rich source of cinnamic acid derivatives so, these markers (cinnamic acid derivatives) can used for routine quality control of Cichorium intybus seed and its extract.

First evidence and characterisation of rare chrome-based colourants used on 19th-century textiles from Myanmar

First evidence for the use of the chrome yellow dyeing method was obtained on late 19th-century Karen textiles from Myanmar. Non-invasive observations obtained by digital microscopy and fibre optic reflectance spectroscopy (FORS) provided hints of the possible presence of non-conventional organic colourants in yellow, orange and green threads used to produce three textiles under investigation. Micro-samples analysed by scanning electron microscopy coupled to energy dispersive X-ray spectroscopy (SEM-EDX) and micro-Raman spectroscopy enabled PbxCrOy formulations directly precipitated onto cotton fibres to be identified and characterised in these threads. Complementary analyses by high pressure liquid chromatography coupled to diode array detector and tandem mass spectrometry (HPLC-DAD-MS/MS) revealed that in the orange and green threads the chrome-based colourants were over-dyed with local natural dyes, thus revealing an unprecedented dyeing scenario, in which cotton threads imported from Europe were over-dyed locally by Karen people to obtain desired colour shades. Additionally, while yellow threads showed the presence of chrome yellow (lead chromate - PbCrO4), orange and green threads mostly contained chrome orange (basic lead chromate – Pb2CrO5 or PbCrO4·Pb(OH)2), which led us to hypothesise that chrome yellow partially transformed into chrome orange due to the over-dyeing process performed in alkaline conditions. This study raises awareness of the need for detailed spectroscopic characterisation coupled with high resolution mass spectrometric analysis to study complex mixtures of dyeing materials in historic textiles and has strong implications for advancing our understanding of the impact of British colonisation on traditional textile-making in Myanmar.

Evolution of anthocyanin content during grape ripening and characterization of the phenolic profile of the resulting wine by comprehensive two-dimensional liquid chromatography

The typical phenolic profile in grapes is characterized by its complexity both in terms of number of diverse chemical structures and their variation during ripening. Besides, the specific phenolic composition of grapes directly influences the presence of those components in the resulting wine. In this contribution, a new method based on the application of comprehensive two-dimensional liquid chromatography coupled to a diode array detector and tandem mass spectrometry has been developed to obtain the typical phenolic profile of Malbec grapes cultivated in Brazil. Moreover, the method has been demonstrated to be useful to study how the phenolic composition in grapes evolved during a 10-week ripening period. Main detected compounds in grapes and in the wine derived from them were anthocyanins, although a good number of polymeric flavan-3-ols were also tentatively identified, among other compounds. Results show how the amount of anthocyanins present in grapes was increased during ripening up to 5–6 weeks and then decreased towards week 9. The two-dimensional approach applied was demonstrated to be useful for the characterization of the complex phenolic profile of these samples, involving more than 40 different structures and has the potential to be further applied to the study of this important fraction is different grapes and wines systematically.

A comprehensive analysis of the chemical composition of Fraxinus mandshurica roots based on HPLC-DAD-MS, GC-MS and UPLC-ESI-Q-TOF-MS

Fraxinus mandshurica is widely distributed around the world, which has significant economic value and pharmacological effects. However, its roots are often overlooked during processing and use. In this study, the preliminary identification of chemical components in F. mandshurica roots was carried out using ultra-performance liquid chromatography coupled with electrospray quadrupole time-of-flight mass spectrometry (UPLC-ESI-Q-TOF-MS), and a total of 37 characteristic components were identified, including 13 secoiridoids, 7 lignans, 6 coumarins, 4 flavonoids, 4 phenylethanols, 1 terpenoid and 2 other compounds. Meanwhile, to evaluate the 6 lignans content of F. mandshurica roots, the identification, quantitation analysis and methodological validation were performed by high-performance liquid chromatography with diode array detection mass spectrometry (HPLC-DAD-MS), which showed that the concentration range of standard compounds was 0.5–1000 μg/mL. The standard curve correlation coefficients (R2) were all greater than 0.9991, indicating that the linearity of the fitted curves was good. The highest lignan content in F. mandshurica roots was olivil (461.11 μg/g) and the lowest in buddlenol E (11.14 μg/g), and the total lignan content was 764.63 μg/g. The relative standard deviations (RSD, %) of both intra-day and inter-day precision were less than 1.95%. The RSD (%) of the reproducibility and stability experiments was less than 2.91%. The spiked recoveries of the samples were in the range of 98.29–102.62%, and the RSD (%) was in the range of 0.43–1.73, indicating that the method has high accuracy. In addition, a total of 20 volatile components in F. mandshurica roots were identified and quantified by gas chromatography-mass spectrometer (GC-MS) in this study, which laid a solid foundation for the comprehensive development and utilization of F. mandshurica resources.

Sterilisation of biopharmaceuticals: Effect of gamma irradiation, e-beam irradiation and nitrogen dioxide on human insulin

Biopharmaceuticals are administered parenterally and therefore sterility is required. Sterility can be obtained via different processes including exposure to steam or dry heat. Sterilisation studies on biopharmaceuticals, which are highly sensitive medicinal products, are scarce. This study investigates the effect of different sterilisation processes on recombinant human insulin in solid state (gamma and e-beam irradiation (w/wo dry ice), nitrogen dioxide (NO2)) and in aqueous solution (gamma irradiation (w/wo dry ice, w/wo glycerin)) using ultra-high performance liquid chromatography-diode array detection-mass spectrometry. It is observed that NO2 substantially degrades the solid samples, while gamma and e-beam irradiation result in lower levels of degradation (mean normalized peak areas of 95.2–96.2 % with respect to the non-sterilised samples). Gamma irradiation of insulin solutions with and without dry ice at 2.5 kGy results in mean normalised peak areas of 85 % and <40 % with respect to the non-sterilised samples, respectively. It is concluded that sterilisation using ionising radiation of liquid biopharmaceuticals with insulin and sterilisation of insulin dry powder using NO2 is less suitable with the set-ups used here because of substantial degradation. In contrast, evidence is presented in favour of sterilisation of insulin dry powder using ionising radiation.

An In Vitro Study of the Healing Potential of Black Mulberry (Morus nigra L.) Extract in a Liposomal Formulation

Natural compounds are used in modern dermal treatments to avoid side effects commonly associated with conventional treatments. The aim of our study was to develop a liposomal formulation including black mulberry extract and to highlight its potential on the healing of normal human dermal fibroblasts (NHDF) in vitro using the scratch test. Mulberry-loaded liposomes (MnL) were prepared using a thin-film hydration method based on cholesterol (C) and phosphatidylcholine (PC) in a 1:3 (w/w) ratio. The liposomal formulation was characterized by analyzing its size, electric surface potential, morphology, entrapment efficiency, and in vitro healing effects. Also, the black mulberry fruits (Morus nigra L.) were characterized from point of view of polyphenolic compounds and antioxidant capacity by Ferric-Reducing Antioxidant Power (FRAP) assay. HPLC-DAD-MS (ESI+) (high performance liquid chromatography-photodiode array detection-mass spectrometry (electrospray ionization)) analysis indicated the presence of phenolic compounds namely from hydroxybenzoic and hydroxycinnamic acids and flavonols. Among flavonols, quercetin-glucoside represented 50.56%, and chlorogenic acid was the predominant compound among hydroxycinnamic acids (37.06%). In vitro fibroblast wound closure was more effective with mulberry-loaded liposomes (MnL) than extracts of mulberries. According to our study, mulberry-loaded liposomes have been shown to be effective in wound healing and can be used as a natural treatment.

Genomic and Metabolite Profiling Reveal a Novel Streptomyces Strain, QHH-9511, from the Qinghai-Tibet Plateau.

The prevalence of superbugs, represented by methicillin-resistant Staphylococcus aureus (MRSA), has become a serious clinical and public safety concern with rising incidence in hospitals. Polyketides with diverse chemical structures harbor many antimicrobial activities, including those of rifampin and rapamycin against MRSA. Streptomyces sp. QHH-9511 was isolated from a niche habitat in the Qinghai-Tibet Plateau and used to produce antibacterial metabolites. Herein, an integrated approach combining genome mining and metabolic analysis were employed to decipher the chemical origin of the antibacterial components with pigmented properties in strain QHH-9511, a novel Streptomyces species from a lichen symbiont on the Qinghai-Tibet Plateau. Genomic phylogeny assembled at the chromosome level revealed its unique evolutionary state. Further genome mining uncovered 36 candidate gene clusters, most of which were uncharacterized. Meanwhile, based on liquid chromatography coupled to diode array detection mass spectrometry, a series of granaticins, BSMs, chromones, phaeochromycins, and related molecules were discovered by using the Global Natural Product Social molecular networking platform. Subsequently, several pigment compounds were isolated and identified by high-resolution mass spectrometry and/or nuclear magnetic resonance, among which the structure-activity relationships of seven aromatic polyketides showed that the fused lactone ring of the C-2 carboxyl group could increase antibacterial activity. Genetic experiments indicated that all seven aromatic polyketides are a series of metabolic shunts produced by a single type II polyketide synthase (PKS) cluster. Comparative genomic analysis of granaticin producers showed that the granaticin gene cluster is widely distributed. This study provides an efficient method to combine genome mining and metabolic profiling techniques to uncover bioactive metabolites derived from specific habitats, while deepening our understanding of aromatic polyketide biosynthesis. IMPORTANCE Undescribed microorganisms from special habitats are being screened for anti-superbug drug molecules. In a project to screen actinomycetes for anti-MRSA activity, we isolated a Streptomyces strain from Qinghai Lake lichens. The phylogeny based on the genome assembled at the chromosome level revealed this strain's unique evolutionary state. The chemical origins of the antibacterial components with pigment properties in strain QHH-9511 were determined using an integrated approach combining genome mining and metabolic analysis. Further genome mining uncovered 36 secondary metabolite gene clusters, the majority of which were previously unknown. A series of aromatic compounds were discovered using molecular network analysis, separation, and extraction. Genetic experiments revealed that all seven aromatic polyketides are a series of metabolic shunts produced by a single cluster of type II PKSs. This study describes a method for identifying novel Streptomyces from specific habitats by combining genome mining with metabolic profiling techniques.

GC-MS and LC-DAD-MS Phytochemical Profiling for Characterization of Three Native Salvia Taxa from Eastern Mediterranean with Antiglycation Properties.

Salvia fruticosa and S. pomifera subsp. calycina are native to Eastern Mediterranean and S. pomifera subsp. pomifera is endemic to Greece. The primary aim of this study was to develop an analytical methodology for metabolomic profiling and to study their efficacy in combating glycation, the major biochemical complication of diabetes. After sequential ultrasound-assisted extraction of 2 g of leaves with petroleum ether and 70% methanol, the volatile metabolites in the petroleum ether extracts were studied with GC-MS (Gas Chromatography-Mass Spectrometry), whereas the polar metabolites in the hydroalcoholic extracts were determined and quantified by UHPLC-DAD-ESI-MS (Ultra-High Performance Liquid Chromatography-Diode Array Detector-Mass Spectrometry). This methodology was applied to five populations belonging to the three native taxa. 1,8-Cineole was the predominant volatile (34.8-39.0%) in S. fruticosa, while S. pomifera had a greater content of α-thujone (19.7-41.0%) and β-thujone (6.0-39.1%). Principal Component Analysis (PCA) analysis of the volatiles could discriminate the different taxa. UHPLC-DAD-ESI-MS demonstrated the presence of 50 compounds, twenty of which were quantified. PCA revealed that not only the taxa but also the populations of S. pomifera subsp. pomifera could be differentiated. All Salvia samples inhibited advanced glycation end-product formation in a bovine serum albumin/2-deoxyribose assay; rosmarinic and carnosic acid shared this activity. This study demonstrates the antiglycation activity of S. fruticosa and S. pomifera extracts for the first time and presents a miniaturized methodology for their metabolomic profiling, which could aid chemotaxonomic studies and serve as a tool for their authentication and quality control.