Spectrometry Method Research Articles

Contaminant Exposure and Liver and Kidney Lesions in North American River Otters in the Indian River Lagoon, Florida.

The harmful algal bloom (HAB) liver toxin microcystin (MC) and trace element biomagnification were previously detected in organisms in the Indian River Lagoon (IRL), Florida. Since there are no routine screening programs for these contaminants, liver tissue from North American river otters (Lontra canadensis), an important sentinel species in the IRL, was screened for MC via enzyme-linked immunoassay (ELISA), followed by confirmatory analyses via liquid-chromatography/mass spectrometry methods (LC-MS/MS). Liver and kidney samples were evaluated for trace element (As, Cd, Co, Cu, Fe, Hg, Mn, Mo, Pb, Se, Tl, and Zn) bioaccumulation via inductively coupled plasma mass spectrometry (ICP-MS). Histopathologic evaluation of the liver and kidney was conducted to assess possible correlation with toxic insults. Forty-three river otters were evaluated (2016-2022). Microcystin was not detected in any river otter sample (n = 37). Of those tested for trace element bioaccumulation (n = 22), no sample measured above provided reference ranges or estimated toxic thresholds for this species. There were no statistically significant patterns observed based on season, year, or age class, but sex had a small influence on trace element levels in the kidney. One individual had a kidney Cu level (52 μg/g dry weight) higher than any previously reported for this species. Trace elements were detected at presumed background levels providing baselines for future monitoring. For otters with available histopathologic evaluation (n = 28), anomalies indicative of contaminant exposure (non-specific inflammation, necrosis, and/or lipidosis) were present in the liver and kidney of 18% and 4% of individuals, respectively. However, since these lesions were not linked to abnormal trace element bioaccumulation or MC exposure, other causes (e.g., infectious disease) should be considered.

Qualitative and quantitative analysis of synthetic cannabinoids in e-liquids by pyrolysis–gas chromatography/mass spectrometry

Synthetic cannabinoids (SCs) are a series of synthetic substances that mimic the effects of natural cannabis and cause more serious toxicity and public safety issues. SCs, with a wide variety and fast update rate have become the most abused family of new psychoactive substances. A pyrolysis–gas chromatography/mass spectrometry method has been developed to qualitatively and quantitatively analyze SCs in e-liquids using pyrolysis products as analytical targets, which can effectively expand the detection range of SCs. In qualitative analysis, diluted e-liquids were pyrolyzed in the microfurnace at 800, 600, and 300 ℃, respectively, followed by separation and analysis through gas chromatography/mass spectrometry. SCs in the e-liquids were determined by both shared and characteristic pyrolysis products combined with the EI mass spectrometry of their parent compounds. ADB-BUTINACA and 5F-MDMB-PICA were successfully identified in five e-liquid samples seized by local police. Furthermore, five popular SCs in the illegal market, ADB-BUTINACA, 4F-ABUTINACA, 4CN-CUMYL-BUTINACA, 4F-MDMB-BICA, and 5F-MDMB-PICA, were quantitatively analyzed based on their characteristic pyrolysis products as the analysis targets. The results indicated that the linear range of ADB-BUTINACA was 2–200 μg/mL, while those of 4F-ABUTINACA and 4CN-CUMYL-BUTINACA were 1–200 μg/mL. For 4F-MDMB-BICA and 5F-MDMB-PICA, the linear range was 5–500 μg/mL. The linear correlation coefficients of determination of five SCs were greater than 0.994. The limit of detection (S/N=3) for five SCs was 0.25–2.50 μg/mL, and the limit of quantification (S/N=10) was 0.50–5.00 μg/mL. This quantitative method was applied to five positive samples in real cases. The results showed that the concentrations of ADB-BUTIANCA and 5F-MDMB-PICA in the positive samples were 751.32–4810.10 μg/mL and 1109.41 μg/mL, respectively. The current research provides a validated pyrolysis–gas chromatography/mass spectrometry method, with simple pre-treatment, fast operation and wide screening range, to accurately identify and quantify SCs in e-liquid samples without reference materials.

Detection Strategies for Volatile Fragrance Released from Agricultural Products: Progress and Prospects

AbstractThe volatile aroma released from agricultural products is closely related to the health status and quality of their growth, thus endowing the related detection with great significance. For example, the dynamic variation of the volatile chemical composition of a banana during the growth process can reflect its ripeness. Also important for quality monitoring and storage is to precisely and swiftly identify volatile compounds produced by mildew in rice and wheat. In this endeavor, the current detection technologies such as gas chromatography‐mass spectrometry method (GC‐MS) cannot meet the pressing needs of smart agriculture in terms of real‐time monitoring, cost‐effectiveness, sensitivity, and detection speed, thereby necessitating alternative strategies to simultaneously satisfy these requirements. Aiming to provide an overall development trend in this field, this paper summarizes the existing detection technologies including GC‐MS, E‐nose, and sensory analysis with their respective shortcomings and challenges, and then proposes the application prospects. This work can effectively enrich the reliable monitoring methods for volatile agricultural fragrance while promoting the long‐run development of smart agriculture.

Non-targeted identification of tianeptine photodegradation products in water samples by UHPLC-QTOF MS/MS

This study aims the characterization of several tianeptine transformation products in ultrapure water by simulated sunlight irradiation. Tianeptine was completely degraded after 106 h of exposition following pseudo-first-order kinetics (half-life time = 12.0 ± 2.4 h). Furthermore, an ultra-high-performance liquid chromatography coupled with a high-resolution quadrupole time-of-flight-mass spectrometry method was developed and fully validated taking into account different method performance parameters for the quantification of tianeptine in river water up to a concentration of 400 pg L−1. Following a non-targeted approach based on mass data-independent acquisition, eight different transformation products not previously reported in the literature were identified and accordingly elucidated, proposing a photodegradation mechanism based on the accurate tandem mass spectrometry information acquired. Irradiation experiments were replicated for a tianeptine solution prepared in a blank river water sample, resulting in the formation of the same transformation products and similar degradation kinetics. In addition, a toxicity assessment of the photoproducts was performed by in silico method, being generally all TPs of comparable toxicity to the precursor except for TP1, and showing a similar persistence in the environment except for TP2 and TP6, while TP4 was the only TP predicted as mutagenic. The developed method was applied for the analysis of four river water samples.

Pharmacokinetic (PK) and pharmacodynamic (PD) findings from a phase 1b study of ATR inhibitor tuvusertib + anti-PD-L1 avelumab in patients with advanced unresectable solid tumors.

2614 Background: Ataxia telangiectasia and Rad3-related protein (ATR) kinase is critical in the DNA damage response, and its inhibition modulates antitumor immunity. The combination of ATR inhibitor (ATRi) + immune checkpoint inhibitor (ICI) has shown activity in patients with ICI-resistant tumors and may have the potential to overcome ICI resistance and induce antitumor immune responses. Methods: Part B1 of the open-label, multicenter study DDRiver Solid Tumors 320 investigated safety, tolerability, PK, and PD, including effects on immune cells, of ATRi tuvusertib in combination with the ICI avelumab (anti-PD-L1) in patients with advanced unresectable solid tumors. Flow cytometry was used to analyze tuvusertib target inhibition via γ-H2AX modulation in the CD45+ fraction of ex-vivo stimulated peripheral blood mononuclear cells, and to explore the effect on the peripheral immunophenotype. Tuvusertib PK samples were analyzed by a validated bioanalytical liquid chromatography/mass spectrometry method. Results: 22 patients were enrolled and treated with tuvusertib 180 mg once daily on a schedule of 2 weeks (w) on treatment followed by a treatment break of 1 or 2 w, and avelumab 800 mg once every 2 weeks (Q2W). The 2 w on/1 w off schedule, corresponding to the recommended dose for expansion (RDE) for tuvusertib monotherapy, was chosen for further exploration. At this schedule, 2 of 9 patients evaluable for dose-limiting toxicity (DLT) experienced DLTs: Grade 3 ALT and Grade 3 AST increase (n=1), and Grade 3 anemia requiring transfusion (n=1). A patient with chordoma experienced a RECIST v1.1 partial response. Preliminary PK data for tuvusertib suggested rapid absorption with median Tmax range of ~2–3 h and mean elimination half-life range of ~2.93 to 4.23 h, with ~2-fold accumulation of steady-state area-under-the-curve following multiple doses. Exposure of tuvusertib in combination with avelumab was consistent with tuvusertib monotherapy exposure (1). PD showed complete or almost complete target inhibition at 1–6 h after tuvusertib 180 mg followed by rebound above baseline after 24 h on days 1 and 8 of cycle 1. No clear trend of variation in absolute counts of myeloid-derived suppressor cells, T and B lymphocytes, monocytes, and natural killer cell subtypes was detected. Conclusions: Tuvusertib and avelumab were combined at established monotherapy doses with no new safety findings. Tuvusertib PD and exposure data were in line with monotherapy observations. The combination did not cause any consistent change of the immunophenotype. Given the accumulating evidence of ATRi as an immunosensitiser (2), further evaluation of this combination in patients with ICI-resistant advanced solid tumors is warranted. 1. Yap T et al., Ann Oncol 2022;33(suppl_7):S197–S224. 2. Besse B et al., JTO 2022;17(suppl_9):S41–S42. Clinical trial information: NCT05396833 .

Pharmacodynamic (PD) and immunophenotyping analyses of ATR inhibitor (ATRi) tuvusertib + ATM inhibitor (ATMi) lartesertib in a phase Ib study in patients with advanced unresectable solid tumors.

2612 Background: Ataxia telangiectasia-mutated (ATM) and Rad3-related (ATR) protein kinases orchestrate the DNA damage response. A synthetic lethal relationship between ATR and ATM genes in cancer has been described1 and ATMi synergistically potentiate the efficacy of ATRi in vitro and in vivo.2 The combination of tuvusertib + lartesertib was investigated in Part A1 of the open-label, multicenter study DDRiver Solid Tumors 320 in patients with advanced unresectable solid tumors. Here, we report results of the PD and immunophenotyping analyses. Methods: PD analysis comprised γ-H2AX in serial blood samples, stimulated ex vivowith 4-NQO, bleomycin, or controls. Flow cytometry was used to measure target inhibition via γ-H2AX modulation in the CD45+ lymphocytes fraction and to explore the effect of tuvusertib + lartesertib on the immunophenotype (myeloid-derived suppressor cells, T and B lymphocytes, monocytes, and natural killer cells subsets) in serial blood samples. Pharmacokinetic samples were analyzed by a validated bioanalytical liquid chromatography/mass spectrometry method. Time-matched blood samples were collected at baseline, 1, 3, 6, and 24 hours after first tuvusertib and lartesertib administration on days 1 and 8 of cycle 1 for the γ-H2AX analysis, and on days 1 and 15 of cycles 1 and 2 before treatment for immunophenotyping. Results: Immunophenotyping data and γ-H2AX levels were obtained from 41 and 34 patients, respectively. For tuvusertib, complete or almost complete target inhibition was seen at 1–6 hours after treatment, followed by a rebound above baseline after 24 hours, on both days 1 and 8 at doses of 130 and 180 mg once daily (QD). For lartesertib, a variable target inhibition of approximately 50 % on average was seen across all time points at doses of 100, 150 and 200 mg QD. No target inhibition was seen for tuvusertib at 90 mg QD and lartesertib at 50 mg QD (cohort 1). Tuvusertib + lartesertib induced a transient decrease of monocytes and natural killer (NK) cells, with partial or complete recovery to baseline levels during treatment breaks in schedules of 2 weeks on treatment followed by a treatment break of 1 or 2 weeks, respectively. Conclusions: Tuvusertib and lartesertib combination PD outcomes were in line with respective monotherapy observations.3,4 The combination did not cause any consistent change in the levels of immune cell subsets at all dose levels tested, except a mild, transient decrease in monocytes and NK cells, in line with the tuvusertib monotherapy observations. 1. Kantidze et al., Trends Cancer 2018;4(11):755–68. 2. Turchick et al., Mol Cancer Ther 2023;22(7):859–72. 3. Yap et al., Ann Oncol 2022;33(suppl_7):S197–S224. 4. Siu et al., Cancer Res 2023;83(8_Suppl):CT171. Clinical trial information: NCT05396833 .

Optimization of microwave‐assisted extraction for quantification of cannabinoids in hemp tea by liquid chromatography‐mass spectrometry

AbstractMicrowave‐assisted extraction (MAE) of cannabinoids from hemp tea was optimized, for the first, using response surface methodology. The effect of temperature (50, 65, and 80°C), irradiation time (4, 7, and 10 min), and solvent‐to‐solid ratio (20, 30, and 40 mL of methanol/g of hemp tea) on cannabinoid extractability were investigated. The concentrations of five cannabinoids, namely Δ9‐tetrahydrocannabinol (Δ9‐THC), cannabidiol (CBD), cannabigerol (CBG), cannabichromene (CBC), and cannabinol (CBN), were selected as response variables. For the quantitative analysis, a liquid chromatography‐mass spectrometry method was developed and validated. The proposed analytical approach demonstrated satisfactory performance characteristics in terms of linearity (R2 ≥ 0.9998), precision (intra‐day: 1.99%–5.97% relative standard deviation [%RSD], inter‐day: 1.95%–6.08%RSD), sensitivity (limit of detection: 1.35–2.36 ng/g, limit of quantification: 4.05–7.08 ng/g) and carry‐over effect (signals ≤ 5.03%), with all cannabinoids eluting within 6 min. For comparison purposes, soxhlet extraction, ultrasound‐assisted extraction (UAE), and conventional‐stirring extraction were additionally performed. MAE proved to be a more effective technique for the extraction of CBD and CBN, while UAE managed to extract Δ9‐THC, CBG, and CBC at higher concentration levels.

The effect of food on the pharmacokinetics of Sutetinib maleate capsule, an irreversible EGFR tyrosine kinase inhibitor, in healthy Chinese subjects.

Sutetinib is an irreversible inhibitor of epidermal growth factor receptor (EGFR) and showed favorable efficacy and safety in patients with locally advanced or metastatic non-small cell lung cancer (NSCLC) harbouring nondrug-resistant rare EGFR mutations. To evaluate the potential food effect, eighteen healthy Chinese subjects were enrolled in a single-centre, randomized, open-label, two-sequence, two-period crossover study. Sutetinib was administered as a single oral 100mg under fasting or fed conditions, and pharmacokinetic sampling was performed following each dose and analysed by a validated liquid chromatography/mass spectrometry method. Safety and tolerability were also evaluated. Food intake slightly decreased maximum plasma concentration (Cmax) and area under the plasma concentration-time curve from time 0 to infinity (AUC0 - inf) of sutetinib (geometric least-squares mean [GLSM] ratio, 80.94% and 86.11%; 90% confidence interval [CI], 68.43-95.72 and 75.88-97.73) and its active metabolite sutetinib N-Oxide (GLSM ratio, 75.58% and 84.00%; 90% CI, 65.69-86.95 and 75.42-93.56), respectively. In addition, the time to maximum plasma concentration (Tmax) of both sutetinib and its metabolite has been prolonged by 2h under fed conditions. A total of 31 adverse events (AEs) occurred during the study, with no serious adverse events (SAE) reported, and no obvious difference was observed between the fasting and fed groups. Our results demonstrated that a high-fat and high-calorie diet caused a significant delay in drug absorption and a marginal reduction in drug exposure. Sutetinib was generally well tolerated in healthy Chinese subjects. (This trial was registered at http://www.chinadrugtrials.org.cn . The registration No. is CTR20201933, and the date of registration is 2020-10-16).

Metastability of Protein Solution Structures in the Absence of a Solvent: Rugged Energy Landscape and Glass-like Behavior.

Native ion mobility/mass spectrometry is well-poised to structurally screen proteomes but characterizes protein structures in the absence of a solvent. This raises long-standing unanswered questions about the biological significance of protein structures identified through ion mobility/mass spectrometry. Using newly developed computational and experimental ion mobility/ion mobility/mass spectrometry methods, we investigate the unfolding of the protein ubiquitin in a solvent-free environment. Our data suggest that the folded, solvent-free ubiquitin observed by ion mobility/mass spectrometry exists in a largely native fold with an intact β-grasp motif and α-helix. The ensemble of folded, solvent-free ubiquitin ions can be partitioned into kinetically stable subpopulations that appear to correspond to the structural heterogeneity of ubiquitin in solution. Time-resolved ion mobility/ion mobility/mass spectrometry measurements show that folded, solvent-free ubiquitin exhibits a strongly stretched-exponential time dependence, which simulations trace to a rugged energy landscape with kinetic traps. Unfolding rate constants are estimated to be approximately 800 to 20,000 times smaller than in the presence of water, effectively quenching the unfolding process on the time scale of typical ion mobility/mass spectrometry measurements. Our proposed unfolding pathway of solvent-free ubiquitin shares substantial characteristics with that established for the presence of solvent, including a polarized transition state with significant native content in the N-terminal β-hairpin and α-helix. Our experimental and computational data suggest that (1) the energy landscape governing the motions of folded, solvent-free proteins is rugged in analogy to that of glassy systems; (2) large-scale protein motions may at least partially be determined by the amino acid sequence of a polypeptide chain; and (3) solvent facilitates, rather than controls, protein motions.

Circulating endogenous sex steroids and risk of differentiated thyroid carcinoma in men and women.

Thyroid cancer (TC) is substantially more common in women than in men, pointing to a possible role of sex steroid hormones. We investigated the association between circulating sex steroid hormones, sex hormone binding globulin (SHBG) and the risk of differentiated TC in men and women within the European Prospective Investigation into Cancer and nutrition (EPIC) cohort. During follow-up, we identified 333 first primary incident cases of differentiated TC (152 in pre/peri-menopausal women, 111 in post-menopausal women, and 70 in men) and 706 cancer-free controls. Women taking exogenous hormones at blood donation were excluded. Plasma concentrations of testosterone, androstenedione, dehydroepiandrosterone, estradiol, estrone and progesterone (in pre-menopausal women only) were performed using liquid chromatography/mass spectrometry method. SHBG concentrations were measured by immunoassay. Odds ratios (ORs) were estimated using conditional logistic regression models adjusted for possible confounders. No significant associations were observed in men and postmenopausal women, while a borderline significant increase in differentiated TC risk was observed with increasing testosterone (adjusted OR T3 vs T1: 1.68, 95% CI: 0.96-2.92, ptrend = .06) and androstenedione concentrations in pre/perimenopausal women (adjusted OR T3 vs T1: 1.78, 95% CI: 0.96-3.30, ptrend = .06, respectively). A borderline decrease in risk was observed for the highest progesterone/estradiol ratio (adjusted OR T3 vs T1: 0.54, 95% CI: 0.28-1.05, ptrend = .07). Overall, our results do not support a major role of circulating sex steroids in the etiology of differentiated TC in post-menopausal women and men but may suggest an involvement of altered sex steroid production in pre-menopausal women.

The Association between the Expression of MicroRNA-4270 and MicroRNA-4441 with some Metabolic Factors in Iranian Rheumatoid Arthritis Patients.

MicroRNAs (miRs) play a role in several diseases, such as rheumatoid arthritis (RA). The purpose of this study was to discover new microRNAs and investigate their involvement in RA, examining their connections with inflammation and metabolic markers. New microRNAs related to RA were predicted using Mirbase and TargetScan databases based on RA target genes. The relationships between miRNAs and targets were visualized with Cytoscape software. Real-time polymerase chain reaction confirmed detectable miRNAs and metabolic factors were assessed using immunoassay and spectrometry methods in RA patients and healthy subjects. Four microRNAs (hsa-miR-153-5p, hsa-miR-4270, hsa-miR-4441, and hsa-miR-6754-5p) showed the highest correlation with RA target genes among millions of microRNAs. The expression of miR-146b (fold change=1.8) and miR-4441 (fold change=1.7) was notably reduced, while miR-4270 showed upregulation (fold change=1.8) in plasma from RA patients compared to healthy individuals. MiR-6754 exhibited a decrease (fold change=1.3) but was statistically insignificant. MiR-153-5p expression was undetectable in plasma. Receiver operating characteristic (ROC) curve analysis indicated that miR-4441, with an area under the ROC curve (AUC) of 0.7728, and miR-4270 (AUC=0.7353) were promising biomarkers for RA. The expression of these studied miRNAs significantly correlated with essential clinical characteristics, including liver enzymes, cholesterol, phosphorus, and vitamin D3. Our findings suggest that miR-4270 and miR-4441, present in the circulation, exhibit distinct expression patterns in RA. These microRNAs may serve as links between inflammation and metabolism and represent promising new biomarkers for this disease.

Natural Radioactivity Levels of the Beach Sands of Cleopatra Beach and Damlatas Beach (Türkiye) and Their Impact on Human Health.

HPGe γ spectrometry method was used to measure the natural radioactivity levels (40K, 232Th, and 226Ra) of the beach sand samples from Cleopatra Beach and Damlatas Beach in Antalya (Türkiye). The mean 40K, 232Th, and 226Ra radioactivity levels of the studied samples were calculated as 276.88 ± 17.24, 25.04 ± 2.88, and 17.06 ± 1.68 Bq/kg, respectively. Being below the radiation limits, these values indicate no risk in terms of public health. Moreover, the radiological risk parameters, such as excess lifetime cancer risk, radium equivalent activity (Raeq), absorbed γ dose rate (D), annual effective dose equivalent (Eaed), γ index (Iγ), internal radiation hazard index (Hin), and external radiation hazard index (Hex), were calculated. The values of all of these parameters were found to be below the internationally accepted radiation limit values. In addition, distribution maps showing the radiological situation of the region were generated although they did not pose a hazard to public health. No results were found in the analysis processes related to artificial radioactivity.

Studying the Association of TKS4 and CD2AP Scaffold Proteins and Their Implications in the Partial Epithelial-Mesenchymal Transition (EMT) Process.

Colon cancer is a leading cause of death worldwide. Identification of new molecular factors governing the invasiveness of colon cancer holds promise in developing screening and targeted therapeutic methods. The Tyrosine Kinase Substrate with four SH3 domains (TKS4) and the CD2-associated protein (CD2AP) have previously been linked to dynamic actin assembly related processes and cancer cell migration, although their co-instructive role during tumor formation remained unknown. Therefore, this study was designed to investigate the TKS4-CD2AP interaction and study the interdependent effect of TKS4/CD2AP on oncogenic events. We identified CD2AP as a novel TKS4 interacting partner via co-immunoprecipitation-mass spectrometry methods. The interaction was validated via Western blot (WB), immunocytochemistry (ICC) and proximity ligation assay (PLA). The binding motif of CD2AP was explored via peptide microarray. To uncover the possible cooperative effects of TKS4 and CD2AP in cell movement and in epithelial-mesenchymal transition (EMT), we performed gene silencing and overexpressing experiments. Our results showed that TKS4 and CD2AP form a scaffolding protein complex and that they can regulate migration and EMT-related pathways in HCT116 colon cancer cells. This is the first study demonstrating the TKS4-CD2AP protein-protein interaction in vitro, their co-localization in intact cells, and their potential interdependent effects on partial-EMT in colon cancer.

Assessment of steroid enzymes action in children and adolescents with obesity

Rising prevalence of obesity has become an important impulse to investigate basic mechanisms involved in regulating the energy balance. It is widely accepted that steroids are potent factors affecting glucose, fat, and protein metabolism. Our study was aimed to analyze differences in the total amount of selected enzymes implicated in steroid metabolism in a group of children suffering from obesity and those with normal weight, further subdivided according to sex and pubertal stage. Data were obtained from 187 Caucasian children and adolescents, including 113 patients (63 girls, 50 boys) with obesity and 74 (34 girls, 40 boys) normal weight volunteers. Standard clinical examinations were performed in both groups. To evaluate the impact of puberty, preadolescent children and those with advanced puberty were assessed separately. Urine steroid excretion profiles were analyzed using gas chromatography/mass spectrometry method. Children with obesity revealed several changes in in the total amount of steroid enzymes as assessed by the relevant metabolite proportions, compared to their norm weight peers. Girls showed a significant increase in the activity of 11βHSD1, while boys demonstrated a relevant elevation in 20αHSD action. Regardless of sex, children with obesity showed an increase in the activity of 5β-reductase + 3αHSD complex and a decrease in the involvement of 11βOH-lase. The effect is attenuated when consider pre- and pubertal subgroups. We hypothesize that changes in the activity levels of selected enzymes may be a compensatory mechanism to limit the glucocorticoid exposure of key target tissues as well as to improve metabolic control and reduce long-term complications of obesity.

Application of the Static Headspace Method as a Tool for Rapid Control of Fennel Seed Tea Vapors

Background:: In recent years, there have been tensions surrounding the accessibility of drugs at pharmacies. This situation has led people to turn to alternative medicine with increased use of plants as medicines. Nevertheless, a good knowledge of the plant's chemical composition is necessary for its proper use due to the presence of toxic products. Objective:: The research objectives are to expand a novel use of the static headspace technique to control the chemical composition of the vapor of fennel seed tea (Foeniculum vulgare Mill.) and also constitute an attempt to develop a targeted qualitative analytical method for quality control and safety assurance of a consumer product. Methods:: This study qualitatively investigated the amount of estragole in fennel herbal tea vapour by successful rehabilitation of the static headspace gas chromatography/mass spectrometry method. In addition, an analysis is conducted on the total phenolic and flavonoid content in both tea and methanol extract. Antioxidant activities of all extracts were measured and compared to Gallic acid. Results:: The fennel seed tea showed total phenolic and flavonoid contents at 187.7 ± 17.1 GAE/100 g and 133.8 ± 9.07 mg CE/100 g, respectively. Fennel seeds tea exhibited good DPPH anti-radical action with an IC50 of 92.38 ± 10.64 mg/g. Even though studies on the total phenols, flavonoid contents, and antioxidant activity of this herbal tea have all yielded positive outcomes, the application of the static headspace method combined with gas chromatography and mass spectrometry indicated the presence of estragole at 37.63 ± 3.77%. This compound was found at 80.67 ± 0.29% in the essential oil. Conclusion:: The application of this method has made it possible to reduce the handling time by eliminating the extraction step and solvent use. The presence of estragole at an alarming level makes it clear that employing plants as medicines must be regulated.

Assessment of cheese frauds, and relevant detection methods: A systematic review.

Dairy products are widely consumed in the world due to their nutritional and functional characteristics. This group of food products are consumed by all age groups due to their health-giving properties. One of these products is cheese which has a high price compared to other dairy products. Because of this, it can be prone to fraud all over the world. Fraud in food products threatens the world's food safety and can cause serious damage to human health. There are many concerns among food authorities in the world about the fraud of food products. FDA, WHO, and the European Commission provide different legislations and definitions for fraud. The purpose of this review is to identify the most susceptible cheese type for fraud and effective methods for evaluating fraud in all types of cheeses. For this, we examined the Web of Science, Scopus, PubMed, and ScienceDirect databases. Mozzarella cheese had the largest share among all cheeses in terms of adulteration due to its many uses. Also, the methods used to evaluate different types of cheese frauds were PCR, Spectrometry, stable isotope, image analysis, electrophoretic, ELISA, sensors, sensory analysis, near-infrared and NMR. The methods that were most used in detecting fraud were PCR and spectrometry methods. Also, the least used method was sensory evaluation.

Rapid and simple identification of trace amounts of sodium azide in beverages and bodily fluids followed by derivatization and liquid chromatography-electrospray ionization tandem mass spectrometry.

Sodium azide (NaN3 ) is a toxic chemical agent to humans by ingestion and inhalation with a growing number of intentional exposures and accidental cases over the last few decades. Due to its low molecular weight and lack of any chromophore, its retention and detection by reverse-phase liquid chromatography-ultraviolet-mass spectrometry methods are a challenging task. To be able to confirm azide exposure, we have developed a method to identify azide in both beverages and bodily fluids. The identification of azide (N3 - ) is based on derivatization with N-(2-(bromomethyl)benzyl)-N,N-diethylethanaminium bromide (CAX-B) at 25°C for 15 min followed by LC/ESI-MS/MS analysis, with no other sample preparation. The azide after derivatization (CAX-N3 ) was stable, retainable by LC and sensitively detected by selected reaction monitoring. The ESI-MS/MS fragmentation of the M+ precursor ion produced characteristic product ions at m/z 118, 100, 91 and 86. The calibration curves for CAX-N3 showed linearity over two orders of magnitude with R2 value of 0.99. Low limits of identification of 0.1-0.5ng/mL were obtained in all investigated matrices (drinking water, tea, orange juice, plasma and urine). Compared with previously reported chromatography-based methods, this method that was based on derivatization and LC/ESI-MS/MS analysis was substantially more sensitive, simpler and faster. The method can be used for forensic investigation to confirm azide exposure from fatal use to much smaller intoxication dose.

Ultra-Fast Mass Spectrometry in Plant Biochemistry: Response of Winter Wheat Proteomics to Pre-Sowing Treatment with Iron Compounds.

In recent years, ultrafast liquid chromatography/mass spectrometry methods have been extensively developed for the use in proteome profiling in biochemical studies. These methods are intended for express monitoring of cell response to biotic stimuli and elucidation of correlation of molecular changes with biological processes and phenotypical changes. New technologies, including the use of nanomaterials, are actively introduced to increase agricultural production. However, this requires complex approbation of new fertilizers and investigation of mechanisms underlying the biotic effects on the germination, growth, and development of plants. The aim of this work was to adapt the method of ultrafast chromatography/mass spectrometry for rapid quantitative profiling of molecular changes in 7-day-old wheat seedlings in response to pre-sowing seed treatment with iron compounds. The used method allows to analyze up to 200 samples per day; its practical value lies in the possibility of express proteomic diagnostics of the biotic action of new treatments, including those intended for agricultural needs. Changes in the regulation of photosynthesis, biosynthesis of chlorophyll and porphyrin- and tetrapyrrole-containing compounds, glycolysis (in shoot tissues), and polysaccharide metabolism (in root tissues) were shown after seed treatment with suspensions containing film-forming polymers (PEG 400, Na-CMC, Na2-EDTA), iron (II, III) nanoparticles, or iron (II) sulfate. Observations at the protein levels were consistent with the results of morphometry, superoxide dismutase activity assay, and microelement analysis of 3-day-old germinated seeds and shoots and roots of 7-day-old seedlings. A characteristic molecular signature involving proteins participating in the regulation of photosynthesis and glycolytic process was suggested as a potential marker of the biotic effects of seed treatment with iron compounds, which will be confirmed in further studies.

An integrated fecal metabolomic based on 1 H-NMR and UPLC-QTOF-MS revealed the preventive mechanism of Gushudan on glucocorticoid-induced osteoporotic rats.

Gushudan (GSD) has the effect of strengthening bones and nourishing kidneys. However, its specific intervention mechanism still remains unclear. In this study, to investigate the pathogenesis of glucocorticoid-induced osteoporosis (GIOP) and the preventive mechanism of GSD on GIOP, fecal metabolomics based on 1 H-NMR and ultra-high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry method was established. The changes in endogenous metabolites and the relevant metabolic pathways in the control group, model group, and GSD treatment group were investigated via multivariate statistical analysis. As a result, a total of 39 differential metabolites were identified. Of these, 22 metabolites, such as L-methionine, guanine, and sphingosine, were newly discovered as differential metabolites of GIOP. Amino acid metabolism, energy metabolism, intestinal flora metabolism, and lipid metabolism were significantly changed in the fecal profiles of GIOP rats, and GSD could play an anti-osteoporosis role by regulating these metabolic pathways. Finally, compared with our previous study of the GSD to prevent kidney yang deficiency syndrome, this study suggested that there were some identical differential metabolites and metabolic pathways. It showed that there was some correlation among the metabolic profiles of the intestine, kidney, and bone in GIOP rats. Therefore, this study offered new insights into the in-depth understanding of the pathogenesis of GIOP and the intervention mechanism of GSD.

Identification, Screening and Antibacterial Mechanism Analysis of Novel Antimicrobial Peptides from Sturgeon (Acipenser ruthenus) Spermary.

Fish is an important source of antimicrobial peptides. This study aimed to identify and screen antibacterial peptides with excellent antibacterial activity derived from sturgeon spermary peptides (SSPs) and to analyze their antibacterial activity and mechanism. Liquid chromatography-mass spectrometry/mass spectrometry methods were used to analyze and identify peptide sequences, computational prediction tool and molecular docking methods were used for virtual screening of antimicrobial peptides, and finally, candidate peptides were synthesized by solid-phase synthesis method. The results demonstrate that SSPs have excellent inhibitory activity against Escherichia coli with an inhibitory rate of 76.46%. Most parts of the SSPs were derived from the sturgeon (Acipenser ruthenus) histones, and the coverage of histone H2B was the highest (45%). Two novel peptides (NDEELNKLM and RSSKRRQ) were obtained by in silico prediction tools and molecular docking, which may interact with the DNA gyrase and dihydrofolate reductase of E. coli by forming salt bridges and hydrogen bonds. Compared to the individual peptides, the antibacterial effect was significantly improved by mixing the two peptides in equal proportions. Two novel peptides change the permeability of the E. coli cell membranes and may exert antimicrobial activity by inhibiting the metabolic process of the nucleic acids.