Electrospray Ionization Tandem Mass Spectrometry Research Articles

Preparation and characterization of ferulic oligosaccharides from different sources by cell-free GH10 and GH11 xylanases

The feruloyl oligosaccharides (FOs) produced by the decomposition of plant hemicellulose have broad potential applications in the food and biomedical areas. FOs were prepared through the specific enzymatic degradation of insoluble dietary fiber from different sources by cell-free GH10 and GH11 xylanases. The cell-free GH10 and GH11 xylanases were obtained by the heterologous expression in Escherichia coli. The enzymatic hydrolysis conditions were optimized as follows: temperature 50 °C, pH 5.5, hydrolysis time 12 h, GH10 xylanase addition 101.74 U, and GH11 xylanase addition 121.60 U. The compositions and structural characterization of wheat bran FOs (WB-FOs), corncob FOs (CC-FOs), and buckwheat straw FOs (BS-FOs) were identified by fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance spectroscopy (NMR), high-performance liquid chromatography (HPLC), and electrospray ionization tandem mass spectrometry (ESI-MS). Degrees of polymerization (DP) of WB-FOs, CC-FOs, and BS-FOs were 3–11, 3–7, and 3–6, respectively. Ultraviolet (UV) radiation was investigated in vitro. The results demonstrated that BS-FOs possessed excellent UV resistance and photostability, followed by effectiveness in WB-FOs and CC-FOs. These results have improved our understanding of the relationship between FOs with different structural types and their UV radiation.

Volatile organic compounds (VOC) metabolites in urine are associated with increased systemic inflammation levels, and smokers are identified as a vulnerable population

BackgroundPrevious studies indicated that exposure to VOCs was linked to increased systemic inflammation levels. However, the dose-response relationships between urine VOCs metabolites and systemic inflammation have not been established, and the key metabolite of the toxic compounds has not been identified. MethodsWe used data in 7007 US adults in the NHANES cycles (2011–2018) across 8 years. Urinary VOC metabolites were measured using ultra-performance liquid chromatography and electrospray tandem mass spectrometry (UPLC-ESI/MSMS). VOC metabolites were adjusted by urinary creatinine level before analysis. Systemic inflammation was assessed by systemic immune-inflammation index (SII) and systemic inflammation response index (SIRI) indices. Generalized linear models, restricted cubic splines (RCS), and weighted quantile sum (WQS) regression were applied to evaluate the associations, exposure-response (E-R) curve and identify the key contributor compound, adjusting for gender, age, race, BMI, marital condition, education level, smoking level, alcohol consumption and physical activity. Smoking status was assessed as an effect modifier. ResultsSignificant and robust positive correlations were found between 8 VOC metabolites and both SII and SIRI. They were N-Acetyl-S-(2-carboxyethyl)-L-cysteine (CEMA), N-Acetyl-S-(2-cyanoethyl)-L-cysteine (CYMA), N-Acetyl-S-(3,4-dihydroxybutyl)-L-cysteine (DHBMA), N-Acetyl-S-(3-hydroxypropyl)-L-cysteine (3HPMA), mandelic acid (MA), N-Acetyl-S-(4-hydroxy-2-butenyl)-L-cysteine (MHBMA3), phenylglyoxylic acid (PGA), and N-Acetyl-S-(3-hydroxypropyl-1-methyl)-L-cysteine (HPMMA). The RCS curves showed J-shaped or exponential shaped E-R relationships for most VOC metabolites. WQS regression found that exposure to the mixture of VOC metabolites was related to increased systemic inflammation, and MA was the key VOC metabolite contributing most to systemic inflammation levels. Smokers exhibited higher levels of urinary VOCs and larger susceptibility to VOC-related increases in SII and SIRI compared to non-smokers. ConclusionThis study demonstrated a strong link between urinary VOC metabolites and increased systemic inflammation, and smokers were more susceptible. Our findings highlighted the significance of reducing VOC exposure to mitigate the inflammation levels, particularly for smokers.

Detection of IDH mutation in glioma by desorption electrospray ionization (DESI) tandem mass spectrometry

Desorption electrospray ionization (DESI) tandem mass spectrometry (MS) is used to assess mutation status of isocitrate dehydrogenase (IDH) in human gliomas. Due to the diffuse nature of gliomas, total gross resection is not normally achieved during surgery, leading to tumor recurrence. The mutation status of IDH has clinical significance due to better prognosis in IDH-mutant patients. The mutant IDH converts alpha-ketoglutaric acid (α-KG) into 2-hydroxyglutarate (2HG), which accumulates abnormally in cells. Immunohistochemical staining (IHC) and genetic testing, the gold standards, are incompatible with intraoperative applications but DESI tandem mass spectrometry (MS/MS) can be used to assess the mutation status of IDH enzyme from tissue intraoperatively. Here, on off-line evaluation is made of the performance of two different types of mass spectrometers in characterization of IDH mutation status. The intensity of 2HG is measured against glutamate (Glu), an intrinsic reference molecule, in both tandem MS measurements. In both cases using DESI clear separation between IDH-mutant (mut) and IDH-wildtype (wt) samples (p < 0.0001) is observed, despite the short analysis time. Due to the higher detection sensitivity, multiple reaction monitoring experiments using a triple quadrupole show slightly better performance compared to product ion MS/MS performed on a simple linear ion trap. Both DESI-MS platforms are capable of providing information on IDH mutation status, which might in future be used at the time of surgery to support decision-making on resection regions, especially at tumor margins.

Systematic Characterisation of the Fragmentation of Flavonoids Using High-Resolution Accurate Mass Electrospray Tandem Mass Spectrometry.

Flavonoids are a class of polyphenolic secondary metabolites found in plants. Due to their ubiquity in our daily dietary intake and their major anti-oxidative, anti-inflammatory and anti-mutagenic activities, they have been a major focus of wide-ranging research for the past two decades. Mass spectrometry combined with liquid chromatography is one of the most popular techniques for the analysis of flavonoids. In this study, high-resolution accurate mass electrospray tandem mass spectrometry was used to study 30 flavonoids in both positive and negative ionisation modes. From the data obtained, common losses were summarised and compiled. Dominating neutral losses were tabulated. The radical loss of CH3· was observed in flavonoids containing methoxy groups and three key diagnostic product ions were identified. These were m/z 153 (indicative of two OH groups on ring A) m/z 167 (indicative of one OH and one methoxy group on ring A) and m/z 151 (a flavanol, with no ketone oxygen but two OH groups on ring A). These will be useful in structural elucidation of unknown flavonoids and flavonoid metabolites. Energy breakdown graphs were utilised to distinguish between three pairs of structural isomers, and to help rationalise proposed fragmentation pathways. Lastly, a competition of loss of CH3· and methane was reported for rhamnetin and isorhamnetin in the negative ion mode for the first time. Proposed fragmentation pathways were given to rationalise the differences in peak intensities for this competitive process.

Composition and Biological Activity of Flavonoid-containing Fractions of an Extract from Gratiola officinalis L.

Gratiola officinalis L. (hedge hyssop), a medicinal plant of the Scrophulariaceae family, has diuretic, purgative, and vermifuge properties. It is used as a herbal tea to treat chronic gastroenteritis, renal colic, jaundice, and intestinal worms. Previously, we have found that an extract from G. officinalis is nontoxic and has antitumor, antioxidant, antimicrobial, antiinflammatory, anticachexic, and other properties. Our aims in this study were to separate the G. officinalis extract into individual fractions, to identify the most biologically active fractions, and to examine the chemical composition of these fractions and their biological activity toward A498 renal carcinoma cells. The G. officinalis extract was fractionated by reversed-phase high-performance liquid chromatography, and each fraction was tested for antitumor activity. The active fractions were characterized by UV-visible electron spectral analysis, circular dichroism analysis, Fourier transform infrared spectroscopy, high-performance liquid chromatography, electrospray ionization tandem mass spectrometry, and nuclear magnetic resonance spectroscopy. Two antitumor-active fractions of a flavonoid nature were isolated and chromatographically purified. On the basis of the nuclear magnetic resonance data, the aglycone fragment of the main component of one fraction was found to be structured as 2-(3,4-dimethoxyphenyl)-7-hydroxychroman-4-one, or 3',4'-dimethoxy-7- hydroxyflavanone. The antitumor effect of the most active fraction containing 7-O-glucoside of apigenin, glycoside 7,3'-di-O-luteolin and trace amounts of eupatilin against renal carcinoma A498 cells was manifested in its cytotoxic, cytostatic, apoptotic and autophagosomal activities. In addition, we found 3-(1-2)-glucoside of soyaspogenol B, which is a pentacyclic triterpenoid in the structure.

Efficient Simultaneous Isolation of Pinostrobin and Panduratin A from Boesenbergia rotunda Using Centrifugal Partition Chromatography.

The bioactive flavonoids pinostrobin (PN) and panduratin A (PA) from Boesenbergia rotunda are essential for research and therapeutic applications. This study introduces an innovative method utilizing ultrasound-assisted extraction with n-hexane pre-treatment, followed by one-step centrifugal partition chromatography (CPC) purification. Extraction efficiency was evaluated using ultra high-performance liquid chromatography (UHPLC), and the isolated compounds were characterized through 1H-NMR and liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), adhering to AOAC validation guidelines. Optimal extraction conditions comprised a particle size of 125 μm, a solid-to-liquid ratio of 1:30 g/mL, and a 10 min extraction time, yielding a crude extract of 6.96 ± 0.07%. Using an n-hexane/MeOH/water (5/3.4/1.6, v/v) solvent system in ascending mode, PN (2.16 mg, 98.78% purity) and PA (0.4 mg, 99.69% purity) were isolated from 67 mg of crude extract within 30 min. This streamlined approach enhances purification efficiency, allowing for faster extraction and higher purity, making it a suitable method for commercial applications.

Lemongrass (Cymbopogon citratus) oil nanoparticle synthesis, characteristic, and evaluation of antibacterial and antifungal effects and the influence on hardness of acrylic resin

Context: Acrylic resin is used in dentistry as a removable denture base. It can cause various pathologies when not properly cleaned. One of the pathologies is denture stomatitis caused by Candida albicans and Streptococcus mutans accumulation on the acrylic resin surface. Therefore, microbial agents such as denture cleansers are needed. Aims: To evaluate the characteristics of lemongrass (Cymbopogon citratus) nanoparticles as a better antibacterial and antifungal herbal ingredient and their relationship with acrylic hardness. Methods: C. citratus oil nanoparticles (LON) were synthesized and analyzed by transmission electron microscopy (TEM). Electrospray ionization tandem mass spectrometry (ESI-MS) analysis was used to analyze the characteristics of LON bioactive components. Minimum inhibitory concentration (MIC) and minimum bacterial concentration (MBC) against C. albicans and S. mutans and mechanical hardness test of acrylic were performed. Results: The LON concentration of MIC and MBC against C. albicans and S. mutans was 25 and 100%, respectively. One-way ANOVA showed no significant difference between groups of LON with different concentrations (p=0.687). A paired t-test showed significant differences in acrylic resin hardness before and after treatment of LON with 100% (p=0.022) and 50% (p=0.021) concentration. There was no significant difference in hardness before and after treatment of other concentrations of LON and chlorhexidine as positive control. Conclusions: LON treatment on acrylic resin decreased the growth of C. albicans and S. mutans without altering the mechanical properties (hardness).

Pharmacokinetic study of iptacopan and its two acyl glucuronide metabolites in monkey plasma by liquid chromatography combined with electrospray ionization tandem mass spectrometry.

In this study, a simple and sensitive liquid chromatography tandem mass spectrometric method was developed and validated for the determination of iptacopan and two acyl glucuronidation metabolites in monkey plasma. The plasma sample was precipitated with acetonitrile and then separated on an Acquity UPLC BEH C18 column (2.1 × 100 mm, 1.7 μm) using 0.1% formic acid and 5 mM ammonium acetate in water and acetonitrile as the mobile phase. The mass spectrometry (MS) detection was performed in positive multiple reactions monitoring (MRM) mode with precursor-to-production transitions. The developed assay was validated over the range of 1-2000 ng/mL for three analytes with correlation coefficient (r) more than 0.99. The validation parameters including accuracy, precision, carryover effect, matrix effect, recovery, and stability were all within the acceptable limits. The validated method has been applied to investigate the pharmacokinetics of iptacopan and its two acyl glucuronidation metabolites in monkey plasma. After intravenous administration, iptacopan showed low clearance (2.75 mL/min/kg) in monkey plasma. After oral administration, the bioavailability was 55.43%. The exposure (AUC0-t) of direct acyl glucuronide (AG) of iptacopan accounts for 9.73% of the iptacopan plasma exposure. The AUC0-t of AG of dealkylated metabolite of iptacopan was present at a lower level, accounting for 0.5% of the iptacopan plasma exposure.

Direct aqueous injection LC−ESI-MS/MS analysis for the determination of 23 EU watch list substances in whole water samples

The Watch List (WL) monitoring program in the European Union (EU) aims to identify and prioritize emerging pollutants in the aquatic environment that may pose risks to human health and the environment. Thus, reliable methods are needed to determine these substances. This study presents the development of a simple, fast, and green method for analysing 23 WL substances (4 antibiotics, 9 azole compounds, 6 other pesticides, and 4 sunscreens), covering a range of medium to very low polarity (log Kow 2.2–6.9), in whole environmental water samples using direct aqueous injection (DAI) in combination with liquid chromatography–electrospray ionization tandem mass spectrometry (LC–ESI-MS/MS). To process the samples, 0.5 mL of acetonitrile is added to 1 mL of raw water, followed by brief shaking. The mixture is then centrifuged to remove sedimented suspended solids before being injected into the LC instrument. A rigorous analytical greenness metric for sample preparation (AGREEprep) assigned this procedure a green score of 0.61. The method showed satisfactory performance characteristics for all analytes under investigation, including limits of quantification meeting the WL requirements, recoveries in the range of 79–120%, and intra-day precision with relative standard deviations in the range of 1.9–20%. The method is suitable for screening and monitoring purposes, as confirmed by the analysis of real samples.

Mediating role of systemic inflammation in the association between volatile organic compounds exposure and periodontitis: NHANES 2011–2014

BackgroundVolatile organic compounds (VOCs) are ubiquitous environmental pollutants which have been suggested to have adverse effects on human health. While the influence of environmental pollutant exposures on periodontitis has attracted elevating attention in recent years, the epidemiological evidence on the association between VOCs exposure and periodontitis was scarce. This study aimed to investigate the potential mediating role of systemic inflammation factors in the complex association between VOCs exposure and periodontitis.MethodsUtilizing data from the National Health and Nutrition Examination Survey (NHANES) 2011–2014, we examined the impacts of VOCs exposure on periodontitis. Concentrations of urinary metabolites of VOCs (mVOCs) were measured using electrospray tandem mass spectrometry to evaluate internal VOCs exposure. Multivariable logistic regression, restricted cubic spline regression (RCS), Bayesian kernel machine regression (BKMR) and Quantile g-computation (QGC) models were performed to investigate the impacts of VOCs exposure on periodontitis. Mediation models were applied to assess the mediated effects of systemic inflammation on the association between mixed VOCs exposure and periodontitis. Besides, we analyzed the association between mixed VOCs exposure and periodontitis in stratified age, gender, and smoking status subgroups.Results1,551 participants were ultimately included for further analyses, of whom 45.20% suffering from periodontitis. Multivariable logistic regression and RCS identified positive associations between single urinary mVOCs and periodontitis (P < 0.05). Notably, BKMR and QGC models suggested that mixed VOCs exposure was significantly associated with periodontitis, with 2-Aminothiazoline-4-carboxylic acid (ATCA) contributing the most (conditional posterior inclusion probability = 0.997). Moreover, systemic inflammation markers (leukocyte and lymphocyte counts) were found to partly mediate the association between VOCs exposure and periodontitis (P < 0.05). No interaction effect was identified between mixed VOCs exposure and periodontitis in age, gender and smoking status subgroups (P > 0.05).ConclusionThis study demonstrated a positive association between VOCs exposure and periodontitis, which was potentially mediated by systemic inflammation factors. Further longitudinal researches are demanded to clarify the underlying mechanisms.

IMPACT OF CYP2D6 POLYMORPHISMS IN PREDUCTING OCCURRENCE OF ADVERSE EFFECT TO TAMOXIFEN AND DEVELOPING RECCURENCE IN ER+ BREAST CANCER PATIENETS:

Objectives: Breast cancer is a major public health in Algeria. Tamoxifen has been approved in the treatment of ER+ breast cancer. Some of negative side effects of tamoxifen are frequently a reason for discontinuation of therapy during treatment, which would otherwise be potentially lifesaving. In the current study, we assessed the association between CYP2D6 polymorphisms and tamoxifen efficacy in Algerian population receiving tamoxifen as adjuvant therapy in ER+ breast cancer. Methods: A total of 76 Algerian hormone receptor-positive premenopausal breast cancer patients with adjuvant tamoxifen treatment were investigated (45.36±6.13). DNA genotyping was performed by TaqMan Open Array technology. Tamoxifen and its metabolites levels were measured by ultra-high-performance liquid chromatography (UHPLC) followed by electro spray tandem mass spectrometry (LC-MS/MS). Results: A significant association between the presence of deficit copy of enzyme activity and development of adverse effect after the commencement of tamoxifen therapy. Low plasma endoxifen were observed in patients categorized as (NM/PM), (IM/ IM), (IM/PM) and (PM/PM). Patients with increased plasma endoxifen concentrations were significantly more likely than patients with reduced or null activity to not report recurrences (P&lt;0.05). We realized that the combination genotypes NM/PM, IM/IM, IM/PM, with PM/PM were more strongly associated with disease recurrence and adverse effects than NM carries of CYP2D6*1 allele (P&lt;0.05). Conclusion: Our results affirm that CYP2D6 polymorphism should be considered in predicting the occurrence of adverse effect fatty liver in women treated with tamoxifen. Thus, alternative treatment can be intended and lifestyle modifications can be implemented

Online restricted access molecularly imprinted solid phase extraction coupled with electrospray ionization-tandem mass spectrometry for determination of mebendazole and albendazole in milk samples

Multifunctional materials, such as restricted access molecularly imprinted polymers covered with bovine serum albumin (RAMIP-BSA), are effective alternatives for sample preparation techniques. This material selectively adsorbs analytes while excluding macromolecules, enhancing the analysis's efficiency. Among analytical techniques, ESI-MS/MS (Electrospray Ionization-Tandem Mass Spectrometry) has successfully identified and quantified various molecules, including trace-level drugs. Therefore, we proposed, for the first time, an integrated online extraction/analysis system that combines the benefits of RAMIP-BSA and ESI-MS/MS for analyzing mebendazole (MBZ) and albendazole (ABZ) in milk samples without the need for chromatographic separation. Initially, a RAMIP selective for MBZ was synthesized using the bulk method with methacrylic acid and glycidyl methacrylate. Then, the polymer was covered with bovine serum albumin. Subsequently, this adsorbent was packed in a small column and coupled with an ESI-MS/MS instrument in an online configuration. Milli-Q water was used as the loading and reconditioning mobile phases, and a solution of formic acid in methanol (1:100 v/v) was employed as the elution phase. The system enabled simultaneous extraction and determination of MBZ and ABZ in milk samples. The method exhibited linearity between 15.0 and 125.0 μg L−1 for MBZ and 10.0 and 125.0 μg L−1 for ABZ (with a correlation coefficient exceeding 0.99). The limits of quantification were 15.0 and 10.0 μg L−1 for MBZ and ABZ, respectively. Good precision and accuracy were achieved. The developed method was used to analyze MBZ and ABZ in real milk samples and proved to be a viable alternative to conventional sample preparation and chromatographic techniques.

Insight into the Coextrusion Mechanism between Whey Protein Isolate and Cysteine.

The disulfide cross-linking sites of whey protein isolate (WPI) coextruded with dissolved cysteine (Cys) at concentrations of 0, 20, 40, 60, 80, and 100 mM were analyzed by liquid chromatography electrospray ionization tandem mass spectrometry (LC/MS/MS) combined with pLink software, and the structure and gel water distribution of WPI during coextrusion (≤50 °C) were also investigated. LC/MS/MS demonstrated that α-La (6) and α-La (120) were the most active sites for intermolecular disulfide cross-linking of α-La. Meanwhile, the molecular weight of protein polymers in coextruded WPI-Cys was the largest at 100 mM Cys, and α-lactalbumin was the main reactant for polymerization from the result of SDS-PAGE and size exclusion chromatography. Additionally, the high concentration of Cys caused the secondary structure of WPI to gradually change from a highly ordered to a disordered structure during coextrusion. In addition, with an increasing concentration of Cys, the free sulfhydryl group of proteins and the binding force to immobilized water gradually increased. Therefore, this work revealed the disulfide cross-linking mechanism between WPI and Cys under low-temperature coextrusion at the molecular level, and the obtained coextruded cross-linked WPI could serve as a novel food ingredient with excellent water-holding capacity for the food industry.

Assessing the stability of daptomycin in icodextrin-based peritoneal dialysis solution.

The stability of antimicrobials in peritonitis during peritoneal dialysis (PD) solutions is a critical factor influencing treatment success. This study investigated the stability of daptomycin (DAP) when combined with icodextrin-based PD solution, by measuring DAP concentrations and observing any structural changes. A dose of DAP (350 mg) was dissolved in 7 mL of saline in a clean bench. The solution was then injected into the large compartment (1,260 mL) of NICOPELIQ® Peritoneal Dialysis Solution and thoroughly mixed. Samples were collected at intervals ranging from 0 to 336 h (7 points in total). The concentration of DAP was quantified using high-performance liquid chromatography (HPLC). The structure of any unidentified peaks was determined using HPLC coupled with electrospray ionization tandem mass spectrometry. DAP maintained 90% of the initial concentration in NICOPELIC® for 72 h at room temperature and 12 h at 37 °C. Unidentified peaks, distinct from DAP, were detected during analysis. Further investigation indicated that these peaks corresponded to anhydrated DAP. The findings from this stability study are expected to enhance the effectiveness of outpatient management and preparation for treating peritonitis using DAP.

Conversion of 5α-Androstane-3α,17β-diol to bis[(4-dimethylamino)phenyl carbamate] derivative for sensitive determination of its rat brain level by LC/ESI-MS/MS.

5α-Androstane-3α,17β-diol (3α,5α-Adiol) is a testosterone-derived neurosteroid and has anxiolytic and analgesic effects via γ-aminobutyric acid type A receptors as with the progesterone-derived neurosteroid, allopregnanolone (AP). Although the psychotropic drug-evoked changes in the brain AP concentration have been intensively studied, those in the brain 3α,5α-Adiol concentration remain poorly understood. One of the causes for this is the limited availability of a validated method for quantifying the brain 3α,5α-Adiol with a sufficient sensitivity and specificity, which is described in this study. To enhance the detectability of 3α,5α-Adiol by electrospray ionization-tandem mass spectrometry (ESI-MS/MS), derivatization with 4-dimethylaminobenzoyl azide was employed. The brain sample was purified by solid-phase extraction and the recovered 3α,5α-Adiol and the deuterated internal standard were derivatized, then measured by liquid chromatography (LC)/ESI-MS/MS with selected reaction monitoring. The derivatized 3α,5α-Adiol, i.e., the bis[(4-dimethylamino)phenyl carbamate] derivative, provided the intense doubly-protonated molecule as the precursor ion, then the specific product ion containing the 3α,5α-Adiol-skeleton by collision-induced dissociation. The detectability of 3α,5α-Adiol was eventually increased 1000-fold by derivatization. Separation of the derivatized 3α,5α-Adiol from its stereoisomers and interfering brain components was achieved using a SunShell Biphenyl column with an isopropyl alcohol-containing mobile phase. A good linearity in the sufficient concentration range, acceptable precision and accuracy, and negligible matrix effect were demonstrated by the validation tests. The animal (rat) study using this method revealed that the brain 3α,5α-Adiol levels were unaffected by the administration of fluoxetine (FLX) and clozapine (CLZ), in contrast to the significant increase of the AP levels. An LC/ESI-MS/MS method capable of quantifying 3α,5α-Adiol in the rat brain using a 20-mg tissue was developed and validated. The brain levels of 3α,5α-Adiol had an entirely different behavior from those of AP due to FLX and CLZ administration.

Dynamic Changes in Flavonoids’ Accumulation Pattern in Tilia miqueliana Flowers at Different Developmental Stages Based on Widely Targeted Metabolomic Analysis

Tilia miqueliana is an endemic species belonging to the genus Tilia L. (Tiliaceae) in China, which is known for its fragrant flowers and nectar, but the dynamic changes in metabolites during its growth and development are still unclear. In this study, the metabolic profiles from T. miqueliana flowers at three developmental stages were detected by performing an ultra-performance liquid chromatography–electrospray ionization–tandem mass spectrometry (UPLC-ESI-MS/MS)-based widely targeted metabolomic analysis. A total of 1138 metabolites were detected, with 288 Differentially Accumulated Metabolites (DAMs) determined, flavonoids accounting for the largest proportion. The trend analysis showed that DAMs present seven distinctive patterns, and subclass 5 obtained the largest amount with continuously increased relative content during flower development. The Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation and enrichment analysis of DAMs showed different overlap and variability in metabolic pathways, indicating different directions of flavonoids’ metabolic flux in the three developmental stages. A correlation network analysis further revealed five core metabolites that played essential roles in flavonoid biosynthesis. This research provides comprehensive insights into the exploitation and utilization of T. miqueliana as well as a scientific basis for phylogenetic studies of the genus Tilia.

Polyphenol composition and antioxidant activity of fermentation combined with enzymatic hydrolysis modified Astragalus membranaceus stems

Astragalus membranaceus (AM) roots are a well-known homologous medicine and food in China. AM stems, which are discarded and not used effectively, also contain many active compounds and exhibit beneficial effects. It has the potential to be explored as antibiotic alternative. Fermentation combined with enzymatic hydrolysis (FEH) is an effective strategy for extracting polyphenol and improving the usage of AM stems. Therefore, in this study, the conditions of FEH and extraction for polyphenol in AM stems were screened. The antioxidant activity of extract from AM stems without and with FEH (AMSE and FAMSE) was evaluated. The metabolite profiles of phenolic acids and flavonoids in AMSE and FAMSE were characterized by ultra-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (UPLC–ESI-MS/MS). The results showed that the highest polyphenol content from AM stems was obtained with cellulase and pectinase (1:1, 2000 U/g), moisture content 43%, fermentation temperature 30 °C, and fermentation time 7 days. Selected extraction conditions of polyphenol were ethanol concentration 50%, ultrasonic power 500 W, extraction temperature 35 °C, and extraction time 40 min. On this condition, compared with AMSE, the polyphenol and flavonoid contents in FAMSE were significantly higher. FAMSE exhibited stronger DPPH, hydroxyl radical scavenging rate and reducing power than AMSE. The relative content of 3-(4-hydroxyphenyl)-propionic acid, dihydroferulic acid, isoferulic acid, 4-hydroxybenzoic acid, 4-hydroxyphenyllactic acid, ferulic acid, vanillic acid, syringic acid, gentisic acid, sinapic acid, apigenin, tricin, acacetin, daidzein, genistein, formononetin, prunetin, pratensein, rhamnocitrin and galangin were significantly upregulated in FAMSE.Graphical

Rapid bromate determination using short-column ion chromatography-mass spectrometry: Application to bromate quantification during ozonation

A streamlined, precise, and dependable method for the concurrent quantification of bromate has been established, employing ion chromatography in conjunction with electrospray ionization tandem mass spectrometry (IC-ESI-MS/MS). In an effort to expedite sample analysis duration, the AG18 short column was utilized for rapid separation of the specified analytes. Utilizing a blend of water and acetonitrile as the mobile phase within an optimized gradient elution setting, it was possible to achieve satisfactory resolution in under 3 minutes. Specifically, the mixing of acetonitrile in water samples improves the sensitivity and immunity of the method. This phenomenon may be attributed to the similar proportions of acetonitrile in the sample and the mobile phase, which effectively reduces the viscosity and surface tension of the droplets during the ESI process. Detection limits and quantification thresholds were 0.14 µg/L and 0.41 µg/L, respectively. Analyte recoveries, when spiked into tap water between 2.5 µg/L to 50 µg/L, ranged from 69.5 % to 116.7 %; precision metrics, measured as intra- and inter-day relative standard deviations, spanned 5.1–7.9 %. The devised approach encountered minimal interference from prevalent water matrix constituents, including chloride (50–200 mg/L), sulfate (25–200 mg/L), bicarbonate/carbonate (1–6 mM), and natural organic matter (1–8 mg/L). In environmental contexts, this methodology effectively tracked the production of bromate in controlled ozonation experiments conducted in the laboratory. Segmented ozone dosing reduced bromate production when the solution contained low concentrations of DOC, which provides an alternative strategy for controlling bromate production.

Measurement of atmospheric amines and aminoamides by column adsorption/extraction and hydrophilic liquid chromatography-electrospray-tandem mass spectrometry.

Sampling and chromatography-mass spectrometry methods were investigated to measure atmospheric amines and aminoamides. Amines and their amide derivatives play significant roles in new particle formation (NPF) in the atmosphere, especially diamines and aminoamides have higher NPF potentials compared to monoamines. For amine sampling, silica gel tube collection and formic acid extraction gave good overall recoveries (>93 ± 8%) for mono-, di-, tri-, tetramines, and aminoamides. Two chromatography methods were subjected to analyze the extracted amines. One involved direct analysis using hydrophilic interaction liquid chromatography with carboxyl or diol group functioned separation column (carboxyl-HILIC or diol-HILIC), and the other utilized derivatization with 4-(N,N-dimethylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (DBD-F) and subsequent reversed-phase chromatography (HPLC). Separated amines were detected by electrospray ionization and tandem mass spectrometry in both cases. DBD-F-HPLC method provided good sensitivity for mono- and all polyamines (limit of detection (LOD) < 4.6nM, relative standard deviation (RSD) for 100nM < 9.2%). However, aminoamides could not be detected by DBD-F-HPLC. Carboxyl-HILIC provided good sensitivities for mono- and diamines and aminoamides (LOD < 1.6nM, RSD < 4.8%). Forest air measurement was performed and data obtained by carboxyl-HILIC and DBD-F-HPLC showed good agreement for 1,3-diaminopropane, 1,4-diaminobutane (putrescine) and 1,5-diaminopentane (cadaverine) (R2 = 0.9215-0.9739, n = 7-14). Carboxyl-HILIC method was the best for the amine analysis, and combination with silica gel tube sampling provides atmospheric monitoring available. The developed method can be used not only to study atmospheric chemistry of diamines and aminoamides but also to analyze flavor/odor of foods, flowers and wastes.

A validated LC-MS/MS method for the simultaneous quantification of iothalamate and hippuran in serum and urine for non-radioactive kidney function assessment

A novel liquid chromatography-tandem mass spectrometry method is described for the quantitative determination of the kidney function markers iothalamate and hippuran in human serum and urine. It is based on protein precipitation with methanol followed by dilution of the supernatant for serum and simple dilution for urine. The polar analytes are chromatographically separated by a 6.5-min gradient on a low-ligand density reversed-phase column; detection is performed by electrospray ionization tandem mass spectrometry in the positive ion mode against stable-isotope labeled internal standards.The results of a thorough method validation show that iothalamate and hippuran can be simultaneously quantified in the concentration ranges 0.500–30.0 ng/mL and 10.0–5000 ng/mL for serum and urine, respectively, with values for CV and absolute bias not exceeding 10 %, and with sufficient stability in all relevant matrices and solvents. The method was successfully applied for the analysis of serum and urine samples of multiple individuals who received both iothalamate and hippuran.