Triple Quadrupole Mass Spectrometry Research Articles

High-throughput glycosaminoglycan extraction and UHPLC-MS/MS quantification in human biofluids.

Glycosaminoglycans (GAGs) are linear, unbranched heteropolysaccharides whose structural complexity determines their function. Accurate quantification of GAGs in biofluids at high throughput is relevant for numerous biomedical applications. However, because of the structural variability of GAGs in biofluids, existing protocols require complex pre-analytical procedures, have limited throughput and lack accuracy. Here, we describe the extraction and quantification of GAGs by using ultra-high-performance liquid chromatography coupled with triple-quadrupole mass spectrometry (UHPLC-MS/MS). Designed for 96-well plates, this method enables the processing of up to 82 study samples per plate, with the remaining 14 wells used for calibrators and controls. Key steps include the enzymatic depolymerization of GAGs, their derivatization with 2-aminoacridone and their quantification via UHPLC-MS/MS. Each plate can be analyzed in a single UHPLC-MS/MS run, offering the quantitative and scalable analysis of 17 disaccharides from chondroitin sulfate, heparan sulfate and hyaluronic acid, with a level of precision and reproducibility sufficient for their use as biomarkers. The procedure from sample thawing to initiating the UHPLC-MS/MS run can be completed in ~1.5 d plus 15 min of MS runtime per sample, and it is structured to fit within ordinary working shifts, thus making it a valuable tool for clinical laboratories seeking high-throughput analysis of GAGs. The protocol requires expertise in UHPLC-MS/MS.

A fragmentation study of disaccharide flavonoid C-glycosides using triple quadrupole mass spectrometry and its application for identification of flavonoid C-glycosides in Odontosoria chinensis.

Flavonoid C-glycosides have a wide range of pharmacological activities. However, there are few mass spectrometric research on C,O-disaccharide flavonoid C-glycosides and di-C,O-saccharide flavonoid C-glycosides. Their low-energy collision-induced dissociation (ESI-CID-MS/MS) fragmentation pattern and differences have not been reported, and the fragment ion library is incomplete. Therefore, it was essential to elucidate the fragmentation patterns of disaccharide flavonoid C-glycosides, which is described in this study. Four disaccharide flavonoid C-glycosides such as vitexin-4″-O-glucoside were analyzed by ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometer (UPLC-MS/MS) using electrospray ionization (ESI) in both positive and negative ion modes. Each ion and its proposed fragmentation pathways of the four disaccharide flavonoid C-glycosides were analyzed comprehensively. Finally, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) and the established fragmentation patterns have been used to identify disaccharide flavonoid C-glycosides in Odontosoria Chinensis. The fragmentation pathways of C,O-disaccharide flavonoid C-glycosides and di-C,O-saccharide flavonoid C-glycosides are similar. They both have the mass spectrometric characteristics of O-glycoside and C-glycoside. Product ions after mixed pathways of neutral fragments such as saccharide ring fragment, O-glycosides, H2O, and CH2O elimination appeared in both types of flavonoid C-glycosides, but their relative abundances are significantly different. According to the established fragmentation patterns, di-C,O-saccharide flavonoid glycosides were also found in Odontosoria chinensis. The fragment ions at m/z 431, 413, 341, 311, 293, and 282 in negative ion mode and m/z 293, 282, 577, 559, 541, 523, 529, and 499 in positive ion mode can serves as the main characteristics for identifying C,O-disaccharide flavonoid C-glycosides and di-C,O-saccharide flavonoid C-glycosides.

Association of joint exposure to organophosphorus flame retardants and phthalate acid esters with gestational diabetes mellitus: a nested case-control study

BackgroundOrganic phosphate flame retardants (OPFRs) and phthalate acid esters (PAEs) are common endocrine-disrupting chemicals that cause metabolic disorders. This study aimed to assess the association between joint exposure to OPFRs and PAEs during early pregnancy in women with gestational diabetes mellitus (GDM).MethodsSeven OPFRs and five PAEs were detected in the urine of 65 GDM patients and 100 controls using gas chromatography-tandem triple quadrupole mass spectrometry (GC-MS). The association of OPFRs and PAEs with GDM was assessed using logistic regression, weighted quantile sum (WQS) regression, and Bayesian kernel machine regression (BKMR) models.ResultsLevels of dibutyl phthalate (DBP), di-2-ethylhexyl phthalate (DEHP), diethyl phthalate (DEP), dimethyl phthalate (DMP), tris (2-butoxyethyl) phosphate (TBEP), tributyl phosphate (TBP), tris (2-chloroethyl) phosphate (TCEP), tris (1,3-dichloro-2-propyl) phosphate (TDCPP), tri-ortho-cresyl phosphate (TOCP), and triphenyl phosphate (TPHP) increased in the GDM group, and the OPFRs and PAEs, except for BBP and TMCP, were associated with GDM in the logistic regression analysis. In the WQS model, the mixture of OPFRs and PAEs was significantly positively associated with GDM (OR = 3.29, 95%CI = 1.27–8.51, P = 0.014), with TDCPP having the highest WQS index weight. BKMR analysis reinforced these results, showing that the overall association of joint exposure to the OPFRs and PAEs with GDM increased at exposure levels of the 55th to 75th percentiles. Independent exposure to TDCPP (OR = 1.42, 95%CI = 1.09–1.86, P = 0.011) and TBEP (OR = 1.29, 95%CI = 1.04–1.60, P = 0.023) were associated with an increased risk of GDM.ConclusionsEnvironmental exposure to OPFRs and PAEs is significantly associated with GDM. These findings provide evidence for the adverse effects of exposure to OPFRs and PAEs on the health of pregnant women.

Fecal tryptophan metabolite profiling in newborns in relation to microbiota and antibiotic treatment

In the first days of life, the newborns’ intestinal microbiota develops simultaneously with the intestinal gut barrier and follows intestinal immunity. The mode of delivery shows significant impact on microbial development and, thus, the initiation of the tryptophan catabolism pathway. Further antibiotics (ATB) treatment of mothers before or during delivery affects the microbial and tryptophan metabolite composition of stool of the caesarean- and vaginal-delivered newborns. The determination of microbiome and levels of tryptophan microbial metabolites in meconium and stool can characterize intestinal colonization of a newborn. From 134 samples from the Central European Longitudinal Studies of Parents and Children: The Next Generation (CELSPAC: TNG) cohort study, 16S rRNA gene sequencing was performed, and microbial tryptophan metabolites were quantified using ultra-high-performance liquid chromatography with triple-quadrupole mass spectrometry. Microbial diversity and concentrations of tryptophan metabolites were significantly higher in stool compared to meconium. Treatment of mothers with ATB before or during delivery affects metabolite composition and microbial diversity in stool of vaginal- and caesarean-delivered newborns. Correlation of microbial and metabolite composition shows significant positive correlations of indol-3-lactic acid, N-acetyl-tryptophan and indol-3-acetic acid with Bifidobacterium, Bacteroides and Peptoclostridium. The positive effect of vaginal delivery on newborns’ microbiome development is degraded when mother is treated with ATB before or during delivery.Key points• Antibiotic treatment diminishes the positive effects of vaginal delivery.• Antibiotic treatment affects metabolite and microbial composition in newborns.• Bifidobacterium and Peptoclostridium could be the producer of indole-lactic acid.

Pharmacokinetics of the Material Basis Compounds of Tianshu Capsule in Treating Migraine

Ethnopharmacological relevanceTianshu capsule (TSC) is a traditional Chinese medicine (TCM) preparation and has significant clinical effect on migraine. The composition characteristics of TSC formula are worth exploring for the treatment of migraine. Aim of the studyIdentify the compounds in vivo after oral administration of TSC, Gastrodin (GAS), ferulic acid, senkyunolide G and senkyunolide I in the blood of healthy and migraine rats were used as representative compounds for time-dependent processes investigation. And we focus on explaining the characteristics of TSC treatment on migraine from a pharmacokinetic perspective. Materials and methodsIn this study, Ultra-high performance liquid chromatography with Orbitrap tandem mass spectrometry (UPLC-Orbitrap-MS/MS) system were used to detect the compounds in rat plasma and brain after oral administration of TSC. A sensitive, selective and reliable ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-TQ-MS/MS) system method was established for the simultaneous quantitative analysis of multi-components and evaluate the pharmacokinetic behavior of four main compounds. ResultsA total of 46 compounds were significantly identified in vivo of rat, including 35 compounds in plasma, 7 compounds only in brain and 4 compounds both in plasma and brain. The quantitative method has been confirmed to be feasible for multi-components content determination, pharmacokinetic parameters indicated that the four compounds absorbed into blood rapidly, and senkyunolide I also cross the blood-brain barrier (BBB) quickly into brain. GAS has relatively high concentrations in plasma, and the parameters AUC0-t, CLz/F displayed significant differences between the normal and migraine rats. And the AUC0-t and Cmax of GAS and ferulic acid exhibited dose-dependent way. Conclusionswe added compounds for the pharmacokinetic study of TSC, providing powerful help for clinical medication in the treatment of migraine.

UHPLC-MS/MS analysis of the influence of paclobutrazol application on Ophiopogon japonicus

The pharmacological effects of Ophiopogon japonicus are influenced by its active substances like saponins and flavonoids, which are crucial for assessing its quality. An ultra-high performance liquid chromatography tandem triple quadrupole mass spectrometry (UHPLC-MS/MS) method was developed for the qualitative and quantitative assessment of 20 saponins and flavonoids in O. japonicus. The average recovery of these compounds ranged from 70% to 130%, with relative standard deviations (RSD) between 0.11% and 16.46%. Planting O. japonicus in Santai County (Sichuan, China) and investigated the impact of 0.6, 1.3, 2.8, 5.6, and 11.3g/L concentrations of paclobutrazol on both root yield and the levels of saponins and flavonoids in O. japonicus. Lower paclobutrazol concentrations (1.3g/L) led to increased levels of saponins and flavonoids in O. japonicus such as ophiopogonin D′, ruscogenin and ophiopogonanone F, with paclobutrazol residues measuring 77.17μg/kg DW. This preliminary insight into the influence of paclobutrazol and production of saponin flavonoids in O. japonicus bears significance for its larger industrial production.

Effect of Fermentation Duration on the Chemical Compounds of Coffea arabica from Ultra Performance Liquid Chromatography–Triple Quadrupole Mass Spectrometry and Gas Chromatography–Mass Spectrometry Analysis During the Washed Processing

Effect of Fermentation Duration on the Chemical Compounds of Coffea arabica from Ultra Performance Liquid Chromatography–Triple Quadrupole Mass Spectrometry and Gas Chromatography–Mass Spectrometry Analysis During the Washed Processing

Mediating effect of oxidative stress on blood pressure elevation in workers exposed to low concentrations of benzene, toluene, and xylene (BTX).

To investigate the mediating effect of oxidative stress on the relationships between low-concentration benzene, toluene, and xylene (BTX) exposure and blood pressure in workers. A cross-sectional study involving 841 workers from a petroleum refining enterprise in Hainan, China, was conducted. Among the workers, 615 workers were exposed to low-concentration BTX, and 216 workers were in the control group. S-phenylmercapturic acid (S-PMA), hippuric acid (HA), and methyl hippuric acid (MHA, including the three isomers 2-MHA, 3-MHA, and 4-MHA) were measured in the urine of workers via high-performance liquid chromatography‒tandem triple quadrupole mass spectrometry to assess the internal BTX burden. Oxidative stress markers, blood pressure, and their correlations were analysed in both the exposed and control groups of workers. Mediation analysis was used to investigate the potential role of oxidative stress in the relationship between BTX exposure and blood pressure. The concentrations of BTX at the sampling points in the enterprise were all below the limits stipulated in China's national occupational health criteria: occupational exposure limits for hazardous agents. With respect to the internal burden of BTX, the concentrations of the benzene metabolite S-PMA, the toluene metabolite HA, and the xylene metabolites 3-MHA and 4-MHA in the urine samples in the exposure group were greater than those in the control group (P < 0.05). The correlation analysis results revealed that the concentration of the benzene metabolite S-PMA in workers' urine was positively correlated with diastolic blood pressure (DBP) (r = 0.265, P < 0.05). Compared with those in the control group, DBP was greater (β = 1.363, 95% CI 0.088 -2.639), serum superoxide dismutase (SOD) activity was lower (β = - 0.037, 95% CI - 0.060 to - 0.013), and the serum malondialdehyde (MDA) concentration was greater (β = 0.066, 95% CI 0.022-0.110) in the exposure group. Partial correlation analysis revealed a positive correlation between DBP and MDA (rs = 0.115, P < 0.01). The results of the mediation analysis indicated that MDA was a complete mediator between low BTX exposure and DBP (P < 0.05). Occupational exposure to low concentrations of BTX elevates blood pressure and oxidative stress among workers. A positive correlation between DBP and MDA was observed, with MDA acting as a complete mediator between low-concentration BTX exposure and DBP elevation.

Processing-induced reduction in dianthrones content and toxicity of Polygonum multiflorum: Insights from ultra-high performance liquid chromatography triple quadrupole mass spectrometry analysis and toxicological assessment.

Polygonum multiflorum-induced liver injury (PM-DILI) has significantly hindered its clinical application and development. This study investigates the variation in content and toxicity of dianthrones, the toxic components of P. multiflorum, during different processing cycles. We employed the ultra-high-performance liquid chromatography triple quadrupole mass spectrometry method to quantify six dianthrones in raw P. multiflorum and formulations processed with a method called nine cycles of steaming and sunning. Additionally, toxicity assessments were conducted using human normal liver cell line L02 and zebrafish embryos. Results indicate a gradual reduction in dianthrones content with increasing processing cycles. Processed formulations exhibited significantly reduced cytotoxicity in L02 cells and hepatotoxicity in zebrafish embryos. Our findings elucidate the relationship between processing cycles and P. multiflorum toxicity, providing theoretical support for its safe use.

A comparative study on the antipyretic effect and underlying mechanisms of different bile-fermented Arisaemas

Ethnopharmacological relevanceCattle bile Arisaema (CBA) and Pig bile Arisaema (PBA) are both processed products fermented from Arisaema erubescens (Wall.) Schott and animal bile, which are recorded in China Pharmacopoeia. Traditionally, bile Arisaema was often used for clearing heat and eliminating phlegm, calming wind and calming panic. Modern pharmacological researches suggest that both two drugs exert an antipyretic effect, while there is lack of the systematical and comparative evidence on underlying mechanism. Aim of the studyTo comprehensively clarify the differences and underlying mechanisms of antipyretic effect of the two drugs. MethodsIn this study, an accurate and reliable detection method based on ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-TQ MS) for comparing the content difference of bile acids from the two drugs was successfully established and applied. Besides, a dry yeast-induced fever rat model was established, and rectal temperature and content of pyrogenic cytokines were conducted to evaluate the antipyretic effect of CBA and PBA. Serum and hypothalamus untargeted metabolomics analysis based on ultra-performance liquid chromatography coupled with quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS/MS) technology were performed for elucidating the changes of metabolic profile. ResultsThe results indicated that CBA and PBA both exerted a significantly antipyretic effect, but CBA showed the characteristic of quicker onset and longer duration than that of PBA. The ELISA and western blotting analysis exhibited that the underlying antipyretic mechanism of the two drugs was closely associated with inhibiting inflammation through regulating TLR4/NF-κB signaling pathway. Moreover, the metabolism pathway analysis revealed that lipid metabolism and amino acid metabolism were greatly disturbed, which showed a certain correlation with antipyretic effect of two drugs. ConclusionCollectively, our results delineate a potential mechanism of two different bile Arisaemas against febrile via regulating metabolic disorders and targeting inhibition of inflammation for the improvement of fever symptom of the body. Notably, our current study suggested that CBA might be a better choice for suppressing fever clinically.

Multiple Reaction Monitoring-Mass Spectrometric Immunoassay Analysis of Parathyroid Hormone Fragments with Vitamin D Deficiency in Patients with Diabetes Mellitus.

Current immunoassay techniques for analyzing clinically relevant parathyroid hormone (PTH) circulating fragments cannot distinguish microheterogeneity among structurally similar molecular species. This hinders the identification of molecular species and the capture of target analyte information. Since structural modifications are important in disease pathways, mass spectrometry can detect, identify, and quantify heterogeneous ligands captured by antibodies. We aimed to create a sensitive and selective multiple reaction monitoring-mass spectrometric immunoassay analysis (MRM-MSIA)-based method for detecting and quantifying PTH fragments or proteoforms for clinical research. Our study established MRM transitions using triple-quadrupole tandem mass spectrometry for the signature peptides of five PTH fragments. This method was validated according to FDA guidelines, employing the mass spectrometric immunoassay (MSIA) protocol to bolster detection selectivity and sensitivity. This validated approach was applied by analyzing samples from type 2 diabetes mellitus (T2DM) patients with and without vitamin D deficiency. We found serum PTH fragments associated with vitamin D deficiency in patients with and without T2DM. We developed and validated the MRM-MSIA technique specifically designed for the detection and quantification (amino acid (aa38-44), (aa45-51), and (aa65-75)) of these fragments associated with vitamin D deficiency and T2DM. This study is the first to accurately quantify plasma PTH fragments using MRM-MSIA, demonstrating its potential for clinical diagnostics.

Free and bound phenolic profiles and antioxidant ability of eleven marine macroalgae from the South China Sea.

Marine macroalgae are of broad interest because of their abundant bioactive phenolic compounds. However, only a few previous studies have focused on bound phenolic compounds. In this study, there were significant differences in total phenolic content, total phlorotannin content, total flavonoid content, and antioxidant ability in free and bound forms, as well as in their bound-to-free ratios, among 11 marine macroalgal species from the South China Sea. Padina gymnospora had the highest total phenolic content of free fractions, and total phlorotannin content, total flavonoid content, and antioxidant activity of free fractions. Sargassum thunbergii had the highest total phlorotannin content of bound fractions, whereas Sargassum oligocystum had the highest total flavonoid content and total phenolic content of bound fractions. Moreover, 15 phenolic acids, 35 flavonoids, 2 stilbenes, 3 bromophenols, and 3 phlorotannins were characterized and quantified using ultra-high-performance liquid chromatography with Xevo triple quadrupole mass spectrometry, and 42 phenolic compounds were reported in the bound fractions of seaweeds for the first time. Among the species, the number and amount of free and bound phenolic compounds varied greatly and the main components were different. Padina gymnospora had the largest total phenolic number, while Turbinaria ornata showed the highest total phenolic amount. Coutaric acid and diosmetin were dominant in Sargassum polycystum, and hinokiflavone was dominant in Caulerpa lentillifera, and cyanidin was dominant in the other seaweeds. Hierarchical cluster analysis was used to divide the seaweed species into seven groups. This study revealed that Padina gymnospora, Sargassum thunbergii, Turbinaria ornata, and Sargassum oligocystum are promising functional food resources.

Site-Specific Stability Evaluation of Antibody-Drug Conjugate in Serum Using a Validated Liquid Chromatography-Mass Spectrometry Method.

Antibody-drug conjugate (ADC) consists of engineered antibodies and cytotoxic drugs linked via a chemical linker, and the stability of ADC plays a crucial role in ensuring its safety and efficacy. The stability of ADC is closely related to the conjugation site; however, no method has been developed to assess the stability of different conjugation sites due to the low response of conjugated peptides. In this study, an integrated strategy was developed and validated to assess the stability of different conjugation sites on ADC in serum. Initial identification of the conjugated peptides of the model drug ado-trastuzumab emtansine (T-DM1) was achieved by the proteomic method. Subsequently, a semiquantitative method for conjugated peptides was established in liquid chromatography-hybrid linear ion trap triple quadrupole mass spectrometry (LC-QTRAP-MS/MS) based on the qualitative information. The pretreatment method of the serum sample was optimized to reduce matrix interference. The method was then validated and applied to evaluate the stability of the conjugation sites on T-DM1. The results highlighted differences in stability among the different conjugation sites on T-DM1. This is the first study to assess the stability of different conjugation sites on the ADC in serum, which will be helpful for the design and screening of ADCs in the early stages of development.

Monitoring of 260 pesticides in extra virgin olive oil and risk assessment for consumers within the framework of the European multiannual control program

The aim of the present research was to develop and validate a robust analytical method for the monitoring of 260 pesticide residues in Extra Virgin Olive Oil (EVOO) expanding the 185 molecules requested by the multiannual control program. The analytical procedure included an ultrasound-assisted liquid-liquid microextraction followed by low-pressure gas chromatography (LP-GC) and ultra-high-performance liquid chromatography (UHPLC), both coupled to triple quadrupole mass spectrometry. Matrix-matched calibration curves showed good linearity with coefficients of determination greater than 0.999. Accuracy values ranged from 65.5 % to 122.3 % and from 61.1 % to 133.3 % for LP-GC and UHPLC, respectively. The recoveries ranged from 14.0 % to 177.9 %.Fifty commercial EVOO, from Italian and EU production, were analyzed to assess pesticide contamination during the 2021–2023 harvesting seasons. The research focused on evaluating consumer risk by assessing both chronic and acute dietary exposure, using the Pesticide Residue Intake Model developed by the EFSA.

Ultra high-performance liquid chromatography tandem mass spectrometry: Development and validation of an analytical method for N2-Ethyl-2’-deoxyguanosine in dried blood spot

In the body, ethanol is metabolized to acetaldehyde by alcohol dehydrogenase and CYP2E1. Acetaldehyde is the main carcinogen that forms DNA adducts, N2-Ethylidene-2'-deoxyguanosine (N2-Ethylidene-dG), which can cause DNA damage and lead to cancer. However, N2-Ethylidene-dG is an unstable form at the nucleoside level and is difficult to measure. So it needs to be reduced using NaBH4 to become N2-Ethyl-2'-deoxyguanosine (N2-Et-dG). This study aims to obtain a selective, sensitive, and validated analytical method to determine N2-Et-dG using ultra-high-performance liquid chromatography-tandem mass spectrometry with allopurinol as the internal standard. The N2-Et-dG analysis was performed using a C18 Acquity® Bridged Ethylene Hybrid column of (1.7 μm, 100 mm × 2.1 mm). The sample matrix used was dried blood spots, and then the DNA sample was extracted using the QIAamp DNA Mini Kit. The optimum analysis conditions were obtained in a combination eluent of 0.1 % acetic acid and methanol (60:40 v/v) with a flow rate of 0.1 mL/min and eluted isocratically for 5 min. Quantification analysis was performed using triple quadrupole mass spectrometry with electrospray ionization in positive mode, with detection at m/z 296.16 > 180.16 for N2-Et-dG and 137.03 > 110.05 for allopurinol, respectively. The validated analytical method follows the 2018 USA Food and Drug Administration guidelines with a linear concentration range of 5–200 ng/mL.

A Liquid Chromatography Coupled to Tandem Mass Spectrometry Method for Simultaneous Estimation of Saxagliptin and Avanafil: Application to Pharmacokinetic Drug‐Drug Interaction in Type 2 Diabetic Rat Model

ABSTRACTErectile dysfunction is a common complication of type 2 diabetes. The combination of saxagliptin and avanafil can effectively manage both conditions in individuals with type 2 diabetes. However, evaluating the potential pharmacokinetic interactions for co‐administration of saxagliptin and avanafil is crucial. Here, we developed a sensitive bioanalytical method for simultaneously quantifying saxagliptin and avanafil in rat plasma using liquid chromatography hyphenated to triple quadrupole mass spectrometry. The chromatographic separation of saxagliptin, avanafil, and irbesartan (internal standard) was achieved on an Aquity BEH C18 column (100 × 2.1 mm, 1.7 µm), using 0.1% formic acid in water and acetonitrile as mobile phase with the gradient elution at a flow rate of 0.2 mL/min. Mass detection was carried out using selective reaction monitoring mode, with the product ions for saxagliptin, avanafil, and irbesartan being 180.2, 375.1, and 207.1 m/z, respectively. The developed liquid chromatography coupled to tandem mass spectrometry method was validated as per the US Food and Drug Administration guidelines in rat plasma. The validation findings confirmed the method's robustness and suitability for accurate quantification of saxagliptin and avanafil in rat plasma with the limit of quantification of 1 ng/mL for both the drugs and were linear in the range of 1–2000 ng/mL. The study found no significant pharmacokinetic interaction between saxagliptin and avanafil, suggesting they can be safely co‐administered to patients with type 2 diabetes.

UPLC‐MS/MS and chemometrics analyses reveal chemical profile and anti‐inflammatory activity of Bienertia cycloptera Bunge fractions

AbstractIntroductionBienertia cycloptera is a species belonging to the Chenopodiaceae family. According to earlier reports, a unique research study on the phytochemistry and biological analysis of that species was conducted.ObjectiveThis study presents an integrated metabolomics investigation combined with multivariate analysis of various extractive fractions of B. cycloptera aerial parts. This study is the first attempt to explore the anti‐inflammatory metabolites from B. cycloptera, showing its significance as a valuable traditional medicine.MethodologyBy comparing retention times, quasi‐molecular ions, and MS/MS fragment ions with databases and literature references, metabolite annotation was accomplished using ultra performance liquid chromatography (UPLC)/triple quadrupole mass spectrometry (MS). Moreover, the effects of the studied samples on the gene expression of the four pro‐inflammatory cytokines (IL‐1β, IL‐6, TNF‐α, and INF‐γ) using polymerase chain reaction (PCR) and comparing their results by those caused by piroxicam were tested to determine their anti‐inflammatory efficacy.ResultsChemical profiling revealed diverse metabolites, with 62 chromatographic peaks identified across various chemical classes. UPLC‐MS/MS of different B. cycloptera fractions unveiled distinct chemical profiles. Results showed distinct chemical compositions in each fraction, with petroleum ether fraction enriched in sterols and fatty acids; methylene chloride fraction in alkaloids, sterols, and cardenolides; ethyl acetate fraction in alkaloids, flavonoids, cardenolides, and phenolic acids; and n‐butanol fraction in flavonoids, alkaloids, and phenolic acids. Multivariate data analysis illustrated clustering patterns among petroleum ether, methylene chloride, ethyl acetate, and n‐butanol fractions. OPLS‐DA models were constructed to discern inter‐class differences, identifying discriminatory metabolites. In vitro cytotoxicity and anti‐inflammatory assays demonstrated the safety and efficacy of B. cycloptera fractions, with significant downregulation of pro‐inflammatory markers. Further analysis revealed specific metabolites associated with anti‐inflammatory effects, such as p‐hydroxybenzoic acid, vanillic acid, tachioside, ferulic acid, staphylionoside D, humilixanthin, bergaptol, vulgaxanthin I, and portulacaxanthin III.ConclusionThe findings of this study provide valuable insights into the chemical composition and bioactivity of B. cycloptera fractions, suggesting their potential as therapeutic agents and warranting further investigation.

Impact of salinity on PAH and halogenated PAH contamination and risks during the pickling of Chinese pickles

In this study, solid-phase extraction coupled with gas chromatography–triple quadrupole mass spectrometry was used for analyzing PAHs and XPAHs in pickles. The concentrations were 43.0±3.9 μg/kg for PAH24 (including EPA PAH 16 and EU PAH15+1) and 0.52±0.07 μg/kg for XPAH18 (including 11 chlorinated PAHs and 7 brominated PAHs) in commercially available xuecai, and 28.3±8.7 μg/kg for PAH24 and 0.25±0.11 μg/kg for XPAH18 in laboratory-pickled radish. Throughout the pickling process, concentrations of PAHs generally decreased, while XPAHs in xuecai and chlorinated PAHs (ClPAHs) in radish initially increased before declining. The elevated ClPAH levels in fermented radish compared to their unfermented counterparts, along with the newly formed 1-chlorinated pyrene in xuecai, highlighted the generation of ClPAHs during pickling. At lower salinity levels, the greater reduction in concentrations of PAHs corresponds with the largest increase in ClPAHs, suggesting potential chlorination of some PAHs into ClPAHs via microbial activity. Although concentrations of ClPAHs rose after pickling, the toxic equivalent quotients of PAH42 (including PAH24 and XPAH18) in pickles after fermentation were lower than those in both unfermented and fermenting samples, indicating that mature pickles did not increase the overall toxicity risk associated with PAHs and XPAHs.

Application of electrospray triple quadrupole mass spectrometry in assessing thalassemia in a population.

Thalassemia is widely prevalent in Asian countries and the Middle East region of the world. While the prevalence of α thal is around 5-20% in the population, the incidence of β thal is around 3-4%. The available data on thalassemia in Indians is based on chromatographic or electrophoretic techniques. The present study demonstrates the utility of mass spectrometry in detecting thalassemia in a small, healthy population from Tiptur, Karnataka. The study included a population of 315 individuals, spanning both children and adults, with ages ranging from 4 to 76 years. EDTA whole blood was collected after obtaining their written consent. The intensities of various globin chains were measured using a triple quadrupole mass spectrometer. The ratio of the peak areas of globin chains (α/β, δ/β (%)) served as biomarkers to detect thalassemia. Thirty random samples of α thalassemia were analyzed using GapPCR. The prevalence of a thal is 21.65% (68/314) based on α/β (< =0.8) and the prevalence of β thal is 3.8% (12/314) based on δ/β (%) (> =11.0). GapPCR detected three heteromutants of α deletion, one mutation in the HBA1 gene, and one a triplication (16.7%, 5/30). One sample was excluded from the analysis due to the presence of a variant. The utility of mass spectrometry for screening thalassemia in a population is demonstrated. The higher occurrence of α-thalassemia in the studied population underscores the importance of screening for thalassemia in regions with reportedly high prevalence rates.

Discriminating Benign from Malignant Lung Diseases Using Plasma Glycosaminoglycans and Cell-Free DNA.

We aimed to investigate the use of free glycosaminoglycan profiles (GAGomes) and cfDNA in plasma to differentiate between lung cancer and benign lung disease, in a cohort of 113 patients initially suspected of lung cancer. GAGomes were analyzed in all samples using the MIRAM® Free Glycosaminoglycan Kit with ultra-high-performance liquid chromatography and electrospray ionization triple quadrupole mass spectrometry. In a subset of samples, cfDNA concentration and NGS-data was available. We detected two GAGome features, 0S chondroitin sulfate (CS), and 4S CS, with cancer-specific changes. Based on the observed GAGome changes, we devised a model to predict lung cancer. The model, named the GAGome score, could detect lung cancer with 41.2% sensitivity (95% CI: 9.2-54.2%) at 96.4% specificity (95% CI: 95.2-100.0%, n = 113). When we combined the GAGome score with a cfDNA-based model, the sensitivity increased from 42.6% (95% CI: 31.7-60.6%, cfDNA alone) to 70.5% (95% CI: 57.4-81.5%) at 95% specificity (95% CI: 75.1-100%, n = 74). Notably, the combined GAGome and cfDNA testing improved the sensitivity, compared to cfDNA alone, especially in ASCL stage I (55.6% vs 11.1%). Our findings show that plasma GAGome profiles can enhance cfDNA testing performance, highlighting the applicability of a multiomics approach in lung cancer diagnostics.