High-performance Liquid Chromatography Electrospray Research Articles

Profiling of 3-Formyl rifamycin (3-FR) from sintered fixed dose combinations (SDCs) of rifampicin and isoniazid by using RP-HPLC, LC-ESI-QTOF and UPLC/MS/MS

The rapid and severe decomposition of rifampicin in the presence of isoniazid (INH) in a gastric acidic pH environment is a well-reported phenomenon. It is important to note that the significant decomposition problem of RIF has an impact on its bioavailability, which becomes a major issue in fixed dose combinations (FDCs). In this degradation process, RIF will undergo acid hydrolysis and form 3-FR, an acid degradation product of RIF. Further, it will react with INH and form an insoluble iso-nicotinoyl hydrazone (HYD). The formed HYD is probably not stable in the acidic condition, so it will further convert into INH and 3-FR. Consequently, the generation of 3-FR in the acidic stomach environment can be a critical factor contributing to a substantial decline in RIF bioavailability and warrants careful consideration. Hence, it is imperative to explore the presence of 3-FR not only in in vitro dissolution samples but also in in vivo plasma samples. Therefore, the current study focused on the chemical analysis of 3-FR using additively manufactured sintered drug combinations (SDCs) to evaluate its formation, release and qualitative absorption in both in vitro and in vivo settings. High-performance liquid chromatography (HPLC) and liquid chromatography-electrospray ionization quadrupole time-of-flight high-resolution mass spectrometry (LC-ESI-QTOF-MS/MS) were employed for analytical investigations. The developed LC/MS-MS method was employed to investigate the presence of 3-FR in the rabbit's plasma following a single oral administration of SDCs. The study revealed the significant presence of 3-FR in both the in vitro and in vivo plasma samples obtained from the SDCs of RIF and INH. This finding indicates the potential of altering the strategy through additive manufacturing of pH-dependent SDCs to address the critical issue of RIF.

Exploring the potential of phenolic and antioxidant compounds in new Rosaceae fruits.

Rosaceae fruits have been used in traditional medicine for the prevention and treatment of diseases. However, these fruits have not extensively been studied regarding their phenolic composition. Thus, this research focuses on the determination of phenolic compounds by high-performance liquid chromatography electrospray ionization time-of-flight mass spectrometry, flavan-3-ols by high-performance liquid chromatography with fluorescence detection, and the antioxidant activity by 2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), and ferric reducing antioxidant power of the fruits of five species of genera Crataegus and Sorbus (Rosaceae). We found a total of 71 phenolic compounds from which 30 were identified in these berries for the first time. Crataegus monogyna and Crataegus laciniata revealed higher total phenolic and flavan-3-ol contents than the other species and the highest antioxidant activities. Therefore, the fruits evaluated have demonstrated to be important sources of bioactive compounds with huge potential for being used in nutraceutical or food scopes. Additional studies could be needed to evaluate the influence of the different production areas on the phenolic content. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Association of phthalate exposure with all-cause mortality across renal function status: The U.S. National Health and Nutrition Examination Survey, 2005–2018

BackgroundWide phthalate exposure has been associated with both declines in renal function and an elevated risk of mortality. Whether phthalate-associated risk of premature mortality differs by renal function status remains unclear. MethodsThis study included 9605 adults from the U.S. National Health and Nutrition Examination Survey. Urinary concentrations of 11 phthalate metabolites were assessed using high-performance liquid chromatography-electrospray ionization tandem mass spectrometry. According to estimated glomerular filtration rate (eGFR), participants were grouped as having normal or modestly declined renal functions, or chronic kidney disease (CKD). Multivariable Cox regression models estimated all-cause mortality associated with phthalate exposure, overall and by renal function status. ResultsOverall, Mono-n-butyl phthalate (MnBP), Mono-benzyl phthalate (MBzP), Mono-(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP) and Mono-(2-ethyl-5-carbox-ypentyl) phthalate (MECPP) were associated with an elevated risk of mortality (P-trend across tertile <0.05). Moreover, significant interactions were observed between eGFR and MEHHP, MEOHP, MECPP, DEHP in the whole population (P for interactions <0.05). After stratification by renal function, total Di (2-ethylhexyl) phthalate (DEHP) was additionally found to be associated with mortality risk in the CKD group (HR = 1.12; 95% CI: 1.01, 1.25). Co-exposure to the 11 phthalate metabolites was associated with a higher risk of all-cause mortality in the CKD (HR = 1.47; 95% CI: 1.18, 1.84) and modestly declined renal function group (HR = 1.25; 95% CI: 1.09, 1.44). ConclusionsThe associations between phthalate exposure and risk of all-cause mortality were primarily observed in CKD patients, reinforcing the need for monitoring phthalate exposure in this patient population.

Discovery of novel neutral glycosphingolipids in cereal crops: rapid profiling using reversed-phased HPLC–ESI–QqTOF with parallel reaction monitoring

This study explores the sphingolipid class of oligohexosylceramides (OHCs), a rarely studied group, in barley (Hordeum vulgare L.) through a new lipidomics approach. Profiling identified 45 OHCs in barley (Hordeum vulgare L.), elucidating their fatty acid (FA), long-chain base (LCB) and sugar residue compositions; and was accomplished by monophasic extraction followed by reverse-phased high performance liquid chromatography electrospray ionisation quadrupole-time-of-flight tandem mass spectrometry (HPLC–ESI–QqTOF–MS/MS) employing parallel reaction monitoring (PRM). Results revealed unknown ceramide species and highlighted distinctive FA and LCB compositions when compared to other sphingolipid classes. Structurally, the OHCs featured predominantly trihydroxy LCBs associated with hydroxylated FAs and oligohexosyl residues consisting of two–five glucose units in a linear 1 → 4 linkage. A survey found OHCs in tissues of major cereal crops while noting their absence in conventional dicot model plants. This study found salinity stress had only minor effects on the OHC profile in barley roots, leaving questions about their precise functions in plant biology unanswered.

Geographical distribution-based differentiation of cultivated Angelica dahurica, exploring the relationship between the secretory tract and the quality

Based on geographical distribution, cultivated Chinese Angelica dahurica has been divided into Angelica dahurica cv. ‘Hangbaizhi’ (HBZ) and Angelica dahurica cv. ‘Qibaizhi’ (QBZ). Long-term geographical isolation has led to significant quality differences between them. The secretory structure in medicinal plants, as a place for accumulating effective constituents and information transmission to the environment, links the environment with the quality of medicinal materials. However, the secretory tract differences between HBZ and QBZ has not been revealed. This study aimed to explore the relationship between the secretory tract and the quality of two kinds of A. dahurica. Root samples were collected at seven development phases. High-Performance Liquid Chromatography (HPLC) and Desorption Electrospray Ionization Mass Spectrometry Imaging (DESI-MSI) were used for the content determination and spatial location of coumarins. Paraffin section was used to observe and localize the root secretory tract. Origin, CaseViewer, and HDI software were used for data analysis and image processing. The results showed that compared to QBZ, HBZ, with better quality, has a larger area of root secretory tracts. Hence, the root secretory tract can be included in the quality evaluation indicators of A. dahurica. Additionally, DESI-MSI technology was used for the first time to elucidate the temporal and spatial distribution of coumarin components in A. dahurica root tissues. This study provides a theoretical basis for the quality evaluation and breeding of improved varieties of A. dahurica and references the DESI-MSI technology used to analyze the metabolic differences of various compounds, including coumarin and volatile oil, in different tissue parts of A. dahurica.

Dose-response formation of N7-(3-benzo[1,3]dioxol-5-yl-2-hydroxypropyl)guanine in liver and urine correlates with micronucleated reticulocyte frequencies in mice administered safrole oxide

Safrole oxide (SAFO), a metabolite of naturally occurring hepatocarcinogen safrole, is implicated in causing DNA adduct formation. Our previous study first detected the most abundant SAFO-induced DNA adduct, N7-(3-benzo[1,3] dioxol-5-yl-2-hydroxypropyl)guanine (N7γ-SAFO-G), in mouse urine using a well-developed isotope-dilution high-performance liquid chromatography-electrospray ionization tandem mass spectrometry (ID-HPLC-ESI-MS/MS) method. This study further elucidated the genotoxic mode of action of SAFO in mice treated with SAFO 30, 60, 90, or 120 mg/kg for 28 days. The ID-HPLC-ESI-MS/MS method detected N7γ-SAFO-G with excellent sensitivity and specificity in mouse liver and urine of SAFO-treated mice. Our data provide the first direct evidence of SAFO-DNA adduct formation in rodent tissues. N7γ-SAFO-G levels in liver were significantly increased by SAFO 120 mg/kg compared with SAFO 30 mg/kg, suggesting rapid spontaneous or enzymatic depurination of N7γ-SAFO-G in tissue DNA. Urinary N7γ-SAFO-G exhibited a sublinear dose response. Moreover, the micronucleated peripheral reticulocyte frequencies increased dose-dependently and significantly correlated with N7γ-SAFO-G levels in liver (r = 0.8647; p < 0.0001) and urine (r = 0.846; p < 0.0001). Our study suggests that safrole-mediated genotoxicity may be caused partly by its metabolic activation to SAFO and that urinary N7γ-SAFO-G may serve as a chemically-specific cancer risk biomarker for safrole exposure.

Profiling of regioisomeric triacylglycerols in pistachio nuts by high-performance liquid chromatography-electrospray ionization mass spectrometry

In this contribution, a non-aqueous reversed-phase liquid chromatographic method coupled to electrospray ionization (ESI) tandem mass spectrometry (LC-MS) is used for the identification of triacylglycerol (TAG) down to the regioisomersíc level in pistachio lipids. Furthermore, for effective profiling of TAG, a two-dimensional (2D) map was employed to identify the entire homologous series of TAGs especially, odd-numbered fatty acid and long-chain derivatives. Notably, TAGs were observed as pairs of regioisomers with distinct retention times and tandem mass spectra. Furthermore, more than 100 TAGs containing 26 fatty acids with 51-63 carbon atoms and 0-3 double bonds were identified in different pistachio samples. The developed method is effective and facile for the fast quantification of complex TAGs in pistachio samples at high selectivity.

Compositions and Sources of Organic Aerosol in PM2.5 in Nanjing in China

Organic aerosols are harmful to the environment because of their impact on air quality and visibility. They have serious effects not only on living beings and ecosystems because of their biological toxicity, but they also have an indirect effect on regional climate change as cloud condensation nuclei and radiation force. Many measures have been applied to decrease air pollution. Although the air quality has greatly improved, the standard of the World Health Organization (WHO) is far from being met at present. In this study, fine particulates were collected in Nanjing throughout 2019, and high-performance liquid chromatography–electrospray ion–mass spectrometry/mass spectrometry (HPLC-ESI-MS/MS) was carried out to determine 14 organic acids, 10 nitrated phenols, 1 aldehyde, and 1 ketone in aerosol samples. In this study, we further determined the changes in the pollutants in Nanjing in recent years compared to previous studies and characterized more kinds of species in the air. We found that different kinds of nitrated phenols showed similar trends of being abundant in winter and substituted in spring, autumn, and summer. 4-Nitrophenol was the most abundant species (2.83 ng m−3) among the nitrated phenols. p-Coumaric acid presented the highest level in summer with an average concentration of 1.55 ng m−3, indicating that grass burning was significant in summer, possibly due to wheat stalk and perennial ryegrass burning. The positive matrix fraction (PMF) model was applied to identify the sources of aerosols in Nanjing, including coal burning, grass burning, softwood burning, hardwood burning, anthropogenic secondary organic aerosols (SOAs), and biogenic SOAs. Coal burning and softwood burning contributed much more to the total determined species with values of 20.3% and 18.2%, respectively. Anthropogenic SOAs contributed 17.1%, and hardwood burning contributed 16.7%. The contribution of biogenic SOAs was 15%, and the grass-burning source contribution was the lowest, with 12.6%. With consideration of the large contribution from anthropogenic combustion activities, more strict measures are required to reduce emission pollutants in the future.

Abscisic Acid: A Potential Secreted Effector Synthesized by Phytophagous Insects for Host-Plant Manipulation.

Abscisic acid (ABA) is an isoprenoid-derived plant signaling molecule involved in a wide variety of plant processes, including facets of growth and development as well as responses to abiotic and biotic stress. ABA had previously been reported in a wide variety of animals, including insects and humans. We used high-performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-(ESI)-MS/MS) to examine concentrations of ABA in 17 species of phytophagous insects, including gall- and non-gall-inducing species from all insect orders with species known to induce plant galls: Thysanoptera, Hemiptera, Lepidoptera, Coleoptera, Diptera, and Hymenoptera. We found ABA in insect species in all six orders, in both gall-inducing and non-gall-inducing species, with no tendency for gall-inducing insects to have higher concentrations. The concentrations of ABA in insects often markedly exceeded those typically found in plants, suggesting it is highly improbable that insects obtain all their ABA from their host plant via consumption and sequestration. As a follow-up, we used immunohistochemistry to determine that ABA localizes to the salivary glands in the larvae of the gall-inducing Eurosta solidaginis (Diptera: Tephritidae). The high concentrations of ABA, combined with its localization to salivary glands, suggest that insects are synthesizing and secreting ABA to manipulate their host plants. The pervasiveness of ABA among both gall- and non-gall-inducing insects and our current knowledge of the role of ABA in plant processes suggest that insects are using ABA to manipulate source-sink mechanisms of nutrient allocation or to suppress host-plant defenses. ABA joins the triumvirate of phytohormones, along with cytokinins (CKs) and indole-3-acetic acid (IAA), that are abundant, widespread, and localized to glandular organs in insects and used to manipulate host plants.

Effect of in vitro Simulated Poultry Digestion on Bioaccessibility of Quercetin 3-D-galactoside and Antioxidants from Moringa oleifera Leaf Extracts

The use of Moringa oleifera leaves as a poultry feed has been recognized worldwide, thanks to its multi-beneficial properties for broiler quality and production. However, research on the absorption and bioaccessibility of antioxidant compounds in this material is still limited. This study aims to investigate the effect of simulated poultry digestion on the bioaccessibility of quercetin 3-D-galactoside (Q3G) from Moringa leaf extracts and its antioxidant capacity via an in vitro model. Initially, a bio-guided fractionation procedure was employed to isolate Q3G and co-occurring antioxidants from Moringa leaves using column chromatography. The identification of Q3G and major phytocompounds within the potent active samples was carried out using high-performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/ MS). The findings revealed that Q3G is predominantly present in the ethyl acetate extract of Moringa leaves, with the highest concentration recorded as 1.3 mg/g of dry weight. Via a simulated poultry digestion model, the bioaccessibility of Q3G in the small intestinal stage was calculated to have the highest proportion of 67.1%. Significantly, the change in Q3G content was in accordance with the tendency for antioxidant activity of the Q3G-included fractions throughout the digestion. The correlation analysis suggested that Q3G has an important role in the antioxidant capacity of Moringa leaf extracts. To the best of our knowledge, this study represents a pioneering attempt to investigate the bioaccessibility and antioxidant activity of Q3G and Q3G-included extracts from Moringa leaves, utilizing an in vitro poultry digestion model. The findings establish a crucial foundation for future research and development endeavors that aim to produce antioxidant supplements enriched with Q3G from natural sources for applications in poultry production.

Separation and identification of terpene-conjugated curcuminoids based on LC-MS/MS and their in vitro anti-inflammatory activities.

Terpene-conjugated curcuminoids are conjugates of curcuminoids and bisabolanes in the rhizomes of Curcuma longa L. The fragmentation pathways of known three terpene-conjugated curcuminoids (bisabolocurcumin-ether, bisabocurcumin and demethoxybisabolocurcumin ether) and curcumin, demethoxycurcumin and bisdemethoxycurcumin were investigated using high-performance liquid chromatography-electrospray ionization tandem mass spectrometry in negative mode to rapidly search and discover similar unknown compounds of the acetone fraction of turmeric. Subsequently, compounds 1-3 were founded in the acetone fraction based on molecular weight and above fragmentation pathways (the characteristic fragment ions, the most and second most abundant fragment ions produced in MS2 spectra). Terpecurcumin X (1) and terpecurcumin Y (3) were further separated by LC-MS/MS guided isolation technique to verify their structures by nuclear magnetic resonance, electrospray ionization high-resolution mass spectroscopy, ultraviolet and visible spectra and infrared spectra. Interestingly, 1 and 3 were new compounds. The results indicate the feasibility and significant advantages of LC-MS/MS for the rapid discovery and analysis of new constituents in traditional Chinese medicine. In vitro, Terpene-conjugated curcuminoids had better nitric oxide inhibitory activity than the other seven curcuminoids (demethoxycurcumin, bisdemethoxycurcumin, curdione, curcumenone, bisacurone, curcumenol, and germacron). This article is protected by copyright. All rights reserved.

Enantiomeric purity analysis of synthetic peptide therapeutics by direct chiral high-performance liquid chromatography-electrospray ionization tandem mass spectrometry

The determination of chiral purity is critical to the evaluation of the quality of peptide pharmaceutical products. For synthetic peptides, the undesirable d-isomers can be introduced as impurities in amino acid starting materials and can also be formed during peptide synthesis and in some cases during product shelf life. A chiral high-performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) method is described that facilitates rapid and accurate determination of amino acid chiral purity of a peptide. The peptide is hydrolyzed in deuterated acid to facilitate correction for any racemization occurring during this step of sample preparation, and the amino acids are subsequently separated by chiral chromatography interfaced with ESI-MS/MS for quantitation. The amino acid samples are analyzed directly following hydrolysis using high-low chromatography and extraction of selected ion response, providing efficiency and simplicity by avoiding the derivatization steps and multiple external standards required by traditional methodologies. GMP method validation feasibility is described for all nineteen chiral proteogenic amino acids. The practical application of the chiral HPLC-ESI-MS/MS method was demonstrated through the recovery of d-amino acid substitutions at each residue of an octapeptide across the 0.1–1.0 % range of interest. The method was applied to the analysis of four model peptides, each consisting of 8–14 amino acid residues, and the results were comparable to those provided by traditional testing methods.

Ionic Liquid-Dispersive Micro-Extraction and Detection by High Performance Liquid Chromatography-Mass Spectrometry for Antifouling Biocides in Water.

A simple analytical method was developed and evaluated for the determination of two antifouling biocides using an ionic liquid-dispersive liquid-liquid micro-extraction (IL-DLLME) and a high-performance liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) analysis. Irgarol 1051 and Sea-Nine 211 were extracted from deionized water, lake water, and seawater using IL 1-hexyl-3-methylimidazolium hexafluorophosphate ([HMIm][PF6]) and ethyl acetate as the extraction solvent and the dispersion solvent. Several factors were considered, including the type and volume of extraction and dispersive solvent, IL amount, sample pH, salt effect, and cooling temperature. The developed method resulted in a recovery range of 78.7-90.3%, with a relative standard deviation (RSD, n = 3) less than 7.5%. The analytes were enriched greater than 40-fold, and the limits of detection (LOD) for two antifouling biocides were 0.01-0.1 μg L-1. The method was effectively applied for the analysis of real samples of freshwater as well as samples of seawater.

Comprehensive profiling of endogenous phytohormones and expression analysis of 1-aminocyclopropane-1-carboxylic acid synthase gene family during fruit development and ripening in octoploid strawberry (Fragaria× ananassa)

The non-climacteric octoploid strawberry (Fragaria × ananassa Duchesne ex Rozier) was used as a model to study its regulation during fruit ripening. High performance liquid chromatography electrospray tandem-mass spectrometry (HPLC-ESI-MS/MS) was employed to profile 28 different endogenous phytohormones in strawberry. These include auxins, cytokinins (CKs), abscisic acid (ABA), ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC), jasmonates, and phenolic compounds salicylic acid (SA), benzoic acid (BzA) and phenylacetic acid (PAA) together with their various metabolic forms that have remained largely unexplored thus far. ABA, ACC and CK N6-(Δ2-isopentenyl)adenine (iP) were found to be associated with ripening while ABA catabolites 9-hydroxy-ABA and phaseic acid mimicked the pattern of climacteric decline at the turning phase of strawberry ripening. The content of other CK forms except iP decreased as fruit ripened, as also that of auxins indole-3-acetic acid (IAA) and oxo-IAA, and of jasmonates. Data presented here also suggest that both the transition and progression of strawberry fruit ripening are associated with N6-(Δ2-isopentenyl)adenosine-5′-monophosphate (iPRMP) → N6-(Δ2-isopentenyl)adenosine (iPR) → iP as the preferred CK metabolic pathway. In contrast, the ethylene precursor ACC was present at higher levels, with its abundance increasing from the onset of ripening to the red ripe stage. Further investigation of ripening-specific ACC accumulation revealed the presence of a large ACC synthase (ACS) encoding gene family in octoploid strawberry that was previously unknown. Seventeen ACS genes were found differentially expressed in fruit tissues, while six of them showed induced expression during strawberry fruit ripening. These data suggest a possible role(s) of ACC, ABA, and iP in strawberry fruit ripening. These data add new dimension to the existing knowledge of the interplay of different endogenous phytohormones in octoploid strawberry, paving the way for further investigation of their individual role(s) in fruit ripening.

Characterization and Comparison of Steroidal Glycosides from Polygonatum Species by High-Performance Liquid Chromatography–Electrospray Ionization Mass Spectrometry

Polygonatum species have been used as traditional medicines and functional foods in Asia and Europe since ancient times. In this study, a fast and simple method based on liquid chromatography coupled with electrospray ionization mass spectrometry (LC-ESI-MS) was developed to systematically analyze and identify the steroidal glycosides in four major Polygonatum species distributed in Japan, including P. odoratum, P. falcatum, P. macranthum, and P. sibiricum. As a result, 31 steroidal glycosides were tentatively identified, including 18 known and 13 previously unreported glycosides. Their structures were identified by the interpretation of chromatographic behavior and ESI-MS fragmentation patterns. The identification of 31 steroidal glycosides was indicative of a common biogenetic pathway in Polygonatum species. Our study disclosed the chemical profiling of steroidal glycosides in the plants of Polygonatum species, which will benefit better phytochemotaxonomical and phytochemical understanding and quality control for their medicinal usage.

Application of a three dimensional polyethyleneimine functionalized graphene oxide aerogel as an adsorbent for the determination of phytohormones in ginseng.

Aerogels have attracted considerable attention in sample pretreatment for their outstanding properties, such as the unique porous structure, large surface area and abundant modifiable active sites. The present research reports a three-dimensional interconnected porous network aerogel (PEI-AGO) manufactured based on graphene oxide (GO), polyethyleneimine (PEI) and agar as basic materials through a vacuum freeze-drying treatment. The PEI-AGO aerogel exhibits great potential as a solid phase extraction adsorbent for the selective purification of six endogenous plant hormones in conjunction with high performance liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS). Several factors affecting the extraction efficiency were investigated. Under the optimized extraction conditions, a wide linear range of 0.5-100 ng mL-1 with a good linearity (r > 0.9934) was observed. Low limits of detection (LODs) and limits of quantification (LOQs) were obtained in the range of 0.032-0.155 ng mL-1 and 0.107-0.518 ng mL-1, respectively. Furthermore, the relative recoveries for spiked ginseng samples exhibited remarkable consistency, ranging from 90.2% to 117.6%, with a relative standard deviation (RSD) of ≤9.4% (n = 3). In summary, PEI-AGO has proven to be an effective adsorbent for the pretreatment and enrichment of phytohormones which can be used for the determination of trace endogenous acidic plant hormones in ginseng leaves.

Analysis of phytoestrogens from eleven soybean cultivars using LC-MS approach

The beneficial effects of soybean isoflavones for the human body have been confirmed, and such as antioxidant activity and suppresses cancer occurrence. Therefore, soybean is widely used as food in the world. In the past few decades, several soybean varieties have been developed. A high-performance liquid chromatography electro-spray ionization with tandem mass spectrometry (HPLC-ESI-MS/MS) approach was developed and validated to determine isoflavone in 11 soybean varieties of Taiwan. The separation of 12 isoflavones was realized with a Phenomenex F5 column (2.1 × 50 mm, 1.7 µm) and using acetonitrile as mobile phase over a total analytical run time of 15 min. An Agilent 6470 triple quadrupole mass spectrometer (QqQ) with multiple reaction monitoring (MRM) was employed in ESI positive mode was achieved for isoflavone analysis. In this method validation includes the linear relationship in the range of 3.0 to 800 ng/mL (R2 > 0.99); extraction recovery (71.2% to 93.6%); matrix effect (71.3% to 84.9%); intra- and inter-day accuracy were within 81 to 118% and intra- and inter-day precision below 15%. The LC-MS approach was successfully achieved to assess isoflavone of 11 soybean varieties, and the average total isoflavone content was 7150 ng/mL. In addition, this approach can contribute to robust applications as a workflow for the stable and sensitive detection of soybean seeds.

Saffron Characterization by a Multidisciplinary Approach

Saffron is a spice obtained from the drying process of the stigmas of the flower Crocus sativus Linnaeus. It is well known that the organoleptic characteristics of this spice are closely linked to the production area and harvesting year. The present work aims to evaluate whether saffron samples produced in different years and origins present sensibly different crocin profiles. To achieve this goal, 120 saffron samples were harvested between 2016 and 2020 in four different Italian areas. The crocins were analysed, identified, and quantified by high-performance liquid chromatography-electrospray-tandem mass spectrometry (HPLC-ESI-MS/MS) in multiple reaction monitoring mode (MRM). Subsequently, ANOVA-simultaneous component analysis (ASCA) was used to evaluate whether the origin and annuity significantly affected the composition of the crocins. ASCA confirmed the relevance of these effects. Eventually, soft independent modelling by class analogy (SIMCA) models were created for each of the four different origins. Mixtures of saffron from different areas were also prepared to test the robustness of the models. SIMCA provided satisfying results; in fact, models provided 100% sensitivity for three origins (Cascia, Sardinia, and Città della Pieve) on the external test set (48 samples) and 88% (sensitivity on the external test set) for the Spoleto class.

Biodegradation of Punicalagin into Ellagic Acid by Selected Probiotic Bacteria: A Study of the Underlying Mechanisms by MS-Based Proteomics.

Pomegranate (Punica granatum L.) is a well-known source of bioactive phenolic compounds such as ellagitannins, anthocyanins, and flavanols. Punicalagin, one of the main constituents of pomegranate, needs to be biodegraded by bacteria to yield metabolites of medicinal interest. In this work, we tested 30 lactic acid bacteria (LAB) and their capacity to transform punicalagin from a punicalagin-rich pomegranate extract into smaller bioactive molecules, namely, ellagic acid and urolithins. These were identified and quantified by high-performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS2). Further, we evaluated the molecular mechanism governing this transformation through label-free comparative MS-based proteomics. All tested LAB strains were capable of transforming punicalagin into ellagic acid, while the biosynthesis of urolithins was not observed. Proteomic analysis revealed an increase of generic transglycosylases that might have a hydrolytic role in the target phenolic molecule, coupled with an increase in the quantity of ATP-binding cassette (ABC) transporters, which might play a relevant role in transporting the resulting byproducts in and out of the cell.

Cranberry Juice Polyphenols Inhibited the Formation of Advanced Glycation End Products in Collagens, Inhibited Advanced Glycation End Product-Induced Collagen Crosslinking, and Cleaved the Formed Crosslinks.

Collagens in the human skin are susceptible to glycation due to their long half-life of about 15 years, accumulating advanced glycation end products (AGEs). The formation of AGEs and the subsequent AGE-induced collagen crosslinking are major factors for skin aging. The objective of this study was to determine the capacity of cranberry juice polyphenols (CJPs) and their fractions to inhibit collagen glycation and to break AGE-induced crosslinks in collagens. Concentrated cranberry juice was extracted to obtain the CJP, which was further fractionated into an ethyl acetate fraction, water fraction, 30% methanol (MeOH) fraction, 60% MeOH fraction, MeOH fraction, and acetone fraction. CJPs and their fractions contained different ratios of anthocyanins, procyanidins, and flavonols. All the fractions significantly inhibited collagen glycation assessed with the collagen-methylglyoxal (MGO) or collagen-dehydroascorbic acid (DHAA) assays. The ethyl acetate fraction and 60% MeOH had the lowest IC50 values in the collagen-MGO and collagen-DHAA assays. The methanol fraction (IC50 = 0.52 μg/mL) and acetone fraction (IC50 = 0.019 mg/mL) had the lowest IC50 values in the inhibition and breakage of AGE-induced collagen crosslinking, respectively. The ethyl acetate fraction significantly scavenged the highest amount of MGO and DHAA after incubation compared to the other fractions. Results suggested that procyanidins were the most effective antiglycation agent in both collagen glycation assays, followed by flavonols and anthocyanins. High-performance liquid chromatography-electrospray ionization─tandem mass spectrometry showed that the reactions of DHAA with quercetin or epicatechin formed several adducts with unreported proposed structures. This study suggested that CJPs may be used as active ingredients in cosmetics to prevent skin collagen glycation and crosslinking and to break the formed crosslinks.