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Viscum coloratum (Kom.) Nakai is a well-known medicinal hemiparasite widely distributed in Asia. The synthesis and accumulation of its metabolites are affected by both environmental factors and the host plants, while the latter of which is usually overlooked. The purpose of this study is to comprehensively evaluate the effects of host and habitat on the metabolites in V. coloratum through multiple chemical and biological approaches. The metabolite profile of V. coloratum harvested from three different host plants in two habitats were determined by multiple chemical method including high-performance liquid chromatography-ultraviolet (HPLC-UV), gas chromatography-flame ionization detector (GC-FID) and ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF/MS). The differences in antioxidant efficacy of V. coloratum were determined based on multiple in vitro models. The multivariate statistical analysis and data fusion strategy were applied to analyze the differences in metabolite profile and antioxidant activity of V. coloratum. Results indicated that the metabolite profile obtained by various chemical approaches was simultaneously affected by hosts and environment factors, and the environment play a key role. Meanwhile, three main differential metabolites between two environment groups were identified. The results of antioxidant assay indicated that environment have more effects on the biological activity of V. coloratum than the host. Therefore, we conclude that the integration of various chemical and biological approaches combined with multivariate statistical and data fusion analysis, which can determine the influences of host plant and habitat on the metabolites, is a powerful strategy to control the quality of semi-parasitic herbal medicine.

The objective of this study was to explore the effects and underlying intervention mechanisms of Angelica water extract (AWE) on abnormal uterine bleeding (AUB) based on serum metabolomics. Firstly, the concentration of main active substances in AWE was determined and the chemical components were identified by UPLC‐Q‐Exactive Orbitrap‐MS/MS. A drug‐induced abortion model was established by mifepristone and misoprostol. After administration AWE (2.16 g/kg) for 7 days, the coagulation function, serum hormone levels, H&E staining, and immunohistochemistry observation of uterus were detected. In addition, serum metabolites profiles were performed on ultra‐performance liquid chromatography–quadrupole–time‐of‐flight mass spectrometry (UPLC‐Q‐TOF‐MS). The contents of ferulic acid, senkyunolide A, and ligustilide in AWE were 0.7276, 0.0868, and 1.9908 mg/g, respectively. Twenty‐six compounds were identified in AWE. It was found that AWE was effective in regulation of coagulation function and promoting endometrial recovery. Meanwhile, the levels of E2, Pg, and HCG and the expression of ERα, Erβ, and PR were down‐regulated in AUB model and up‐regulated by the treatment of AWE. Twenty‐one potential biomarkers were eventually identified by multivariate statistical analysis. Study indicated that glycerophospholipid, sphingolipid, amino acids, retinol metabolism and primary bile acid biosynthesis were the main related metabolic pathways involved for the treatment of AUB by AWE. The results showed that AWE has potential therapeutic effect on AUB by altering the metabolic aberrations., Angelica water extract could reduce alleviate pathological injury and regulate the coagulation function to promote recovery of the endometrium. Angelica water extract could increase the levels of E2, Pg, and HCG and the expression of ERα, Erβ, and PR. Angelica water extract could regulate the metabolic pathways which include the glycerophospholipid metabolism, sphingolipid metabolism, glycine, serine and threonine metabolism, and retinol metabolism.

Background:Udenafil, a recently discovered drug used for erectile dysfunction treatment, has been widely prescribed and its effect on human systems has been extensively studied. However, there is little research on the human metabolites of udenafil. Three metabolites have been identified in rats. Objective: Herein, highly sensitive and accurate liquid chromatography–quadrupole time-of-flight tandem mass spectrometry (LC-Q-TOF-MS/MS) was conducted to identify new udenafil metabolites. Methods:Human liver microsomes were incubated with udenafil for in vitro samples, and rat urine and faeces samples were collected from udenafil-administered rats for in vivo samples. Each sample was deproteinated with acetonitrile and extracted by solid phase extraction. The purified samples were separated and analyzed by LC-Q-TOF-MS, and some metabolite candidates were reanalyzed for further structural analysis using LC-Q-TOF-MS/MS. Results:Eleven and three metabolites were identified in the in vitro and in vivo samples, respectively, and were found to be hydrolyzed, oxidized, or demethylated forms of udenafil or its metabolites. The error of the metabolic analysis was −8.7 to 7.6 ppm, indicating the high accuracy of the method. Conclusion:These metabolic results could be useful for further investigation of udenafil and new phosphodiesterase-5 inhibitors.

Two-dimensional gas chromatography (2D GC)���quadrupole time-of-flight mass spectrometry (QTOFMS) combined with derivatization was applied to determine naphthenic acids (NAs) in oil sand processed water (OSPW). Using the excellent resolution of 2D GC and mass resolution of QTOFMS with derivatization, 8 cyclohexane carboxylic acid isomers, 18 methylcyclohexyl carboxylic acid isomers, and 18 methylcyclohexyl acetic acid isomers were identified. Linear relationships across wide concentration ranges were established with correlation coefficients (R2) of 0.9995, 0.9992, and 0.9990, for cyclohexanecarboxylic acid, 4-methylcyclohexanecarboxylic acid, and 4-methylcyclohexane acetic acid. The precision, expressed as the relative standard deviation (RSD, n = 5), was from 7. 7���9.5% with recoveries between 72.0% and 84.9%. This method was applied to analyze sewage and showed high concentrations of naphthenic acids in untreated samples.

Anabolic-androgenic steroids (AAS) are synthetic derivatives of testosterone which are used medically for several diseases. However, misuse is commonly observed by athletes to promote enhancement of strength and performance. AAS are frequently obtained through online black markets from clandestine drug manufacturing laboratories, without any quality standards, being potentially dangerous for users. The purpose of this work was the development and application of a fast and simple procedure for the quantitation of stanozolol by liquid chromatography coupled to quadrupole time-of-flight-mass spectrometry (LC-QTOF-MS) in tablets packs seized in Rio Grande do Sul state, Brazil. The samples of stanozolol were separated considering its dosage form. The internal standard (methyltestosterone) was added to the aliquots of the samples, dissolved in methanol and 5μL were injected into the analytical system. The newly developed method has been validated for lower limit of quantitation (LLOQ), linearity, accuracy, precision and selectivity. The LLOQ was 0.1 µg/mL. The developed method was successfully applied to 31 samples seized by the Secretaria da Receita Federal do Brasil (a Brazilian federal revenue service agency). According to the results, 90.3% of the suspected medicines (n=31) were adulterated, and 65% exhibited higher concentrations of stanozolol than those indicated in the label. This work successfully established a new method for quantification of stanozolol using LC-QTOF-MS. This method aims at contributing to the identification and quantification of this anabolic androgenic steroid frequently seized by federal inspection agencies.

A liquid chromatography−quadrupole time-of-flight mass spectrometry (LC−QTOF/MS) based plant metabolomics approach was established to investigate the metabolic profiles of the Cinnamomum camphora leaves of four different areas. The LC−QTOF/MS data were subjected to principal component analysis (PCA) and orthogonal partial least squared discrimination analysis (OPLS-DA) to find the potential characteristic components of C. camphora leaves from the four areas in a quick way. C. camphora leaves from different areas could be separated into four different groups chemically according to the PCA scores. The chemical constituents in C. camphora leaves from four areas were obviously different. The potential markers were identified by comparison with the reference compounds or were tentatively assigned by matching the retention time, empirical molecular formula and fragment ions with those of the published compounds of C. camphora leaves. The proposed analytical method is fast, accurate and reliable for differentiating C. camphora leaves from different areas. Moreover, this study supplied a new method for the quality evaluation of other medicinal materials.

Gedi (Abelmoschus Manihot L.) is a tropical plant in the Malvaceae family, one of the groups of plants in the herbal community. Gedi plants are known to contain flavonoids, hormones, alkaloids, tannins and phenolic compounds. An ultrasonic extraction tool is used in the extraction process of Gedi leaves. The objective of the study is to know the yield and analyze Gedi leaf extract, including proximate analysis and active substance components with LCMS-QTof. The material used is Gedi leaves from Manado. Ultrasonic extraction was carried out with a dry Gedi leaf ratio and Ethanol 1:40 (m/v) with a time of 30 minutes. The concentration of the extract is with a rotary evaporator temperature of 50-60℃. The yield of the extract of Gedi leaf is 9.86%. Study of the proximity of dried Gedi leaf powder revealed 27.135% protein, 41.675% carbohydrate, 329 kcal energy, 11.235% water and 13.83% ash material. Meanwhile for proximity products, the Gedi leaf extract contains 0.635% protein, 1.27% carbohydrate, 9.26% total energy, 7.1% water, and 0.635% ash. The active compounds in the Gedi extract have been analyzed using LCMS. Trigonelline, Gentiatibetine, Harman, Periplocoside C, and Biotin were detected using the positive-ionization mode, whereas Chloramphenicol was detected with negative-ionization mode.

The rhizomes of Dioscorea septemloba (DS) is one of traditional Chinese medicine, which can be used for treatments of urethral and renal infections, as well as rheumatism. Nevertheless, detailed information on DS chemical compositions is still scarce. Therefore, the present work is aimed at investigating the phytochemical components of DS using high performance liquid chromatography coupled to electrospray ionization and quadrupole time-of-flight mass spectrometry in negative ion mode. The proposed method provided tentative identification of 32 components, 6 of which were determined to be new. The present study highlighted the importance of DS as a promising source of medicinal ingredients and provides valuable information on the new potential components in DS.

The lucrative market of herbal remedies spurs rampant adulteration, particularly with pharmaceutical drugs and their unapproved analogues. A comprehensive screening strategy is, therefore, warranted to detect these adulterants and, accordingly, to safeguard public health. This study uses the data-dependent acquisition of liquid chromatography-quadrupole time-of-flight-mass spectrometry (LC-QTOF-MS) to screen phosphodiesterase 5 (PDE5) inhibitors in herbal remedies using suspected-target and non-targeted strategies. For the suspected-target screening, we used a library comprising 95 PDE5 inhibitors. For the non-targeted screening, we adopted top-down and bottom-up approaches to flag novel PDE5 inhibitor analogues based on common fragmentation patterns. LC-QTOF-MS was optimised and validated for capsule and tablet dosage forms using 23 target analytes, selected to represent different groups of PDE5 inhibitors. The method exhibited excellent specificity and linearity with limit of detection and limit of quantification of

Three different vegetable oils, including soybean, corn, and sunflower oils, were differentiated from olive oil by using ultra-performance convergence chromatography coupled with quadrupole time-of-flight (UPC2-QTOF MS) and multivariate data analysis based on their differences in triacylglycerol compositions. Then, olive oil was adulterated by adding these three vegetable oils in 1%, 0.75%, and 0.5% (v/v), and the adulterated olive oils were differentiated from the pure olive oils using the similar analytical strategies but different data processing approaches. After that, the representative markers in differentiating the adulterations were selected, and a mathematical model was created to detect the olive oil adulteration based on these specific markers. These results indicated that UPC2-QTOF MS coupled with multivariate data analysis is a sensitive and accurate method in detecting olive oil adulteration, even in 0.5% adulteration level (v/v). This method could be applied in olive oil adulteration detection, and potentially beneficial to the oil industry.

The dried root of Asparagus cochinchinensis (RAC) has been used as an important traditional Chinese medicine for a long time in China. Steroidal saponins (SSs) are considered to be the main active ingredients of this herb. However, the isolation and structural determination of SSs from RAC are time-consuming and laborious. For this reason, the development of new methods for the separation and characterization of SSs is highly desirable. In this study, a new high-performance liquid chromatography coupled to electrospray ionization and quadrupole time-of-flight mass spectrometry (HPLC-ESI-QTOF-MS/MS) method with precursor ions and the corresponding fragment ions was developed for the identification of SSs in RAC. Finally, 30 SSs have been detected and identified, including 17 potential new compounds. This is the first systematic study of SSs in RAC by HPLC-ESI-QTOF-MS/MS method.

Donkey milk has been widely shown to be an ideal substitute for human milk because of its similar composition. However, alterations to the composition of donkey milk during lactation have not been well studied. In this study, untargeted metabolomics with ultra-high-performance liquid tandem chromatography quadrupole time-of-flight mass spectrometry were used to analyze and compare the metabolites in donkey colostrum (DC) and mature milk (DMM). Two hundred seventy metabolites were characterized in both DC and DMM. Fifty-two of the metabolites in the DC were significantly different from those in the DMM; 8 were downregulated and 44 were upregulated. This demonstrated that the composition of the donkey milk changed with lactation. Additionally, the interactions and metabolic pathways were further analyzed to explore the mechanisms that altered the milk during lactation. Our results provide comprehensive insights into the alterations in donkey milk during lactation. The results will aid in future investigations into the nutrition of donkey milk and provide practical information for the dairy industry.

The purpose of this research was to develop a simple, sensitive, and accurate method for simultaneous determination of 35 free amino acids using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Tea samples were extracted with boiling water bath, and then separated by XBridge BEH Amide column by gradient elution. The exact mass and MS/MS spectra of the target compound was detected under the TOF–MS and Information dependent acquisition (IDA)–MS/MS mode. The results demonstrated good linearity (R2 > 0.9980) in the range of 0.5–1,000 ng/mL. The limits of detection (LODs) were 0.13–25.00 mg/kg and the limits of quantitation (LOQs) were 0.25–50.00 mg/kg. The recovery rate ranged from 70.1 to 105.1% with relative standard deviations (RSDs) n = 6). This research provides a targeted strategy for developing an analysis method for amino acids in tea.

Per- and polyfluoroalkyl substances (PFAS) represent a large group of synthetic organic compounds which exhibit unique properties and have been extensively used for consumer and industrial products, resulting in a widespread presence in the environment. Regulation requiring PFAS monitoring has been implemented worldwide due to their potential health and eco-toxicological effects. Targeted methods are commonly used to monitor between twenty to forty PFAS compounds, representing only a small fraction of the number of compounds that may be present. Consequently, there is an increasing interest in complementary non-targeted methods to screen and identify unknown PFAS compounds with the aim to improve knowledge and to generate more accurate models regarding their environmental mobility and persistence. This work details the development of a method that simultaneously provided targeted and non-targeted PFAS analysis. Ultra-high performance liquid chromatography (UHPLC) was coupled to ion mobility-quadrupole time of flight-mass spectrometry (IMS-QTOF-MS) and used to quantify known and screen unknown PFAS in environmental samples collected within the greater Sydney basin (Australia). The method was validated for the quantification of 14 sulfonate-based PFAS, and a non-targeted data analysis workflow was developed using a combination of mass defect analysis with common fragment and neutral loss filtering to identify fluorine-containing species. The optimised method was applied to the environmental samples and enabled the determination of 3-7 compounds from the targeted list and the detection of a further 56-107 untargeted PFAS. This simultaneous analysis reduces the complexity of multiple analyses, and allows for greater interrogation of the full PFAS load in environmental samples.

Purpose The present study aims to develop a forensic toxicological library to identify 56 natural toxic substances by liquid chromatography–quadrupole time-of-flight tandem mass spectrometry (LC–QTOF-MS/MS). Methods For setting up the library of product ion spectra, individual substances (31 plant toxins, 7 mushroom toxins, 5 marine toxins, 5 frog venoms, 4 mycotoxins, and 4 substances derived from plants) were analyzed by LC–QTOF-MS/MS with positive and negative ionization. The product ion spectra were acquired at the collision energies (CEs) of 20, 35, and 50 eV in single enhanced product ion mode and then in collision energy spread mode in which the CE ramp range was set to 35 ± 15 eV. Results To test the performance of the library, human blood plasma samples were spiked with a mixture of lycorine and domoic acid, extracted by acetonitrile deproteinization and analyzed by LC–QTOF-MS/MS. Identification by our library search could be achieved for these toxins at the purity scores of 79.1 and 67.2, respectively. The method was also applied to postmortem blood from a death case with an aconite intake, and showed that four toxins in an aconite could be identified in the blood sample at the purity scores of 54.6–60.3. Conclusions This library will be more effective for the screening of natural toxic substances in routine forensic toxicological analysis. To our knowledge, there are no reports dealing with development of library for natural toxic substances by LC–QTOF-MS/MS.

The whole plant of Plantago depressa is commonly used as traditional Chinese medicine and functional food ingredient. Total phenolic content and total flavonoid content evaluation indicated that its ethyl acetate and n-butanol fractions were rich in polyphenols. However, to date, the polyphenolic profile is limited because of the complex matrix. Here, polyphenols in ethyl acetate and n-butanol fractions of P. depressa were first separated and enriched by High-Speed Countercurrent Chromatography (HSCCC). Then, their structures were characterized or tentatively identified by High-Performance Liquid Chromatography (HPLC)–Diode Array Detector (DAD)–Quadrupole Time-of-Flight Tandem Mass Spectrometry (QTOF-MS/MS) (HSCCC × HPLC–DAD–QTOF-MS/MS). Because of the orthogonality of HSCCC and HPLC, 37 polyphenols were identified. Notably, 29 polyphenols were discovered from P. depressa for the first time. Finally, contents of two potential new compounds, 2-ethoxy plantamajoside isomers (17 and 29), were determined as 0.27 ± 0.03 and 0.04 ± 0.01 mg/g, respectively. Results indicated that P. depressa could be used as a promising source of polyphenols for health promotion, and the developed method (HSCCC × HPLC) was efficient to explore polyphenolic profile for complex natural products.

Synthetic cannabinoids pose significant threats to public health and safety, as their implications in overdose and adverse events continue to arise in United States and around the world. Synthetic cannabinoids have seen several generations of chemically diverse structural elements, impacting potency and effects. These factors create new analytical challenges for forensic laboratories. This report describes an efficient liquid chromatography/quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) assay for the identification of synthetic cannabinoid parent compounds and metabolites, including real-time identification of emergent compounds, using a SCIEX TripleTOF® 5600+ with non-targeted SWATH® acquisition. Method validation evaluated precision/accuracy, limits of detection, interferences, processed sample stability and carryover, for which 19 parent compounds and 19 metabolites were tested. To demonstrate feasibility, de-identified blood sample extracts were acquired from a large forensic toxicology laboratory and analyzed using the validated LC-QTOF-MS assay. In mid-2018, 200 blood extracts were analyzed, demonstrating a 19% positivity rate with > 94% agreement rate with original testing. In addition, three newly discovered synthetic cannabinoids were identified, including 5F-MDMB-PICA, 4-cyano CUMYL-BUTINACA and 5F-EDMB-PINACA. These synthetic cannabinoids were previously unreported in forensic toxicology casework in the United States. 5F-MDMB-PICA has become the most prevalent synthetic cannabinoid in United States, as of early 2019. These results demonstrate the effectiveness of this assay and workflow in the identification and characterization of synthetic cannabinoids, as well as the usefulness of sample-mining using non-targeted mass acquisition by LC-QTOF-MS for the discovery of NPS. High resolution mass spectrometry should be considered when developing new or novel assays for synthetic cannabinoids.

Branched fatty chain fatty acids (BCFAs) are associated with the "mutton flavor" found with the aroma resulting from cooked older sheep meat with three BCFAs, 4-methyloctanoic (MOA), 4-ethyloctanoic (EOA) and 4-methylnonanoic (MNA) acids as the main compounds responsible for "mutton flavor". Usually, BCFA analysis is done by gas chromatography (GC) with the use of quadrupole mass spectrometry (qMS) becoming predominant. 2-Butyloctanoic acid (2BO) has been used in this facility using as an internal standard to determine BCFA content in sheep fat. In this present work, GC-qMS, along with GC-quadrupole-time of flight MS (GC-QTOF-MS), have been deployed to characterize alkyl esters (as methyl, ethyl, propyl, and butyl) for MOA, EOA, MNA, and 2BO. This work presents, for the first time, the mass spectral characterization of 2BO for these alkyl esters using GC-qMS and GC-QTOF-MS.

Objectives: The study was conducted to identify the phenolic compounds and other possible bioactive compounds present in the leaf extracts of Tabernaemontana heyneana Wall. Methods: Phenolic acid profiling was carried out using ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight (QTOF). An internal standard syringic acid was used for quantitation of phenolic acids and naringenin for quantitation of flavonoids. Results: The leaf extracts analysis revealed the presence of 17 compounds consisting of 14 phenolic compounds and three terpenes. Among 17 compounds, eight were the major compounds, namely, coniferyaldehyde, resveratrol, sinapic alcohol, protocatechuic acid, 4-hydroxybenzaldehyde, chlorogenic acid, rutin, and protocatechuic aldehyde. This forms the first report on the identification of these pharmaceutically important compounds in T. heyneana. Conclusion: These findings offer clear evidence and scientific support for further research on the leaf extract of T. heyneana plant for its therapeutic purpose.

Nitrogen-containing compounds are important components in human breath. However, their origins have not yet been clearly understood. In this study, a modified electrospray ionization (ESI) source coupling with quadrupole time-of-flight mass spectrometry has been used for breath analysis. Fourteen nitrogen-containing compounds were identified in mouth-exhaled breath, and 10 of them were from the oral cavity and oropharynx. Moreover, 8 of these nitrogen-containing compounds were recognized as endogenous metabolites. This result provides important clues for exploring the biological origins of these nitrogen-containing compounds. Observation of the ion suppression phenomenon also indicates that breath analysis should be carried out after clearing of the oral cavity and oropharynx, or directly through nose-breathing to eliminate the influence of those nitrogen-containing compounds from the oral cavity.

Minor lipids in cereals (such as phytosterols and alkylresorcinols) can be important for human nutrition and/or be used as biomarkers for cereal intake. However, the analysis of cereal lipids is very challenging due to the complex lipidome comprising several hundred individual compounds present over a wide range of concentrations. Here we present a method for the profiling of cereal lipids using high temperature gas chromatography coupled to high resolution mass spectrometry (GC/Q-TOF MS). The method was used to investigate the lipid profiles of 77 samples of bread wheat, spelt, einkorn, emmer, barley, rye and oats. Distinct differences in the patterns of alkylresorcinols, free and conjugated sterols and tocopherols between the cereals could be observed. Furthermore, traces of tocomonoenols and diunsaturated and methyl-alkylresorcinols (not previously reported in cereals) could be detected. Finally, the lipid patterns in the cereals could be used to separate the cereals by Principal Component Analysis.

Rice bean (Vigna umbellata) is consumed as a common vegetable in Southeast Asia. Methyl jasmonate (MeJA) functions as an endogenous/exogenous stress signal to elicit the production of specific metabolites during sprouting. To elucidate the changes in metabolites and the effects of MeJA on rice bean sprouting, non-targeted ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry along with principal component analysis and supervised orthogonal projection to latent structure-discriminant analysis was performed to analyze the dissimilar profiles among the sprouts at different stages. Analysis of the abundance of biomarkers, such as sugars, free fatty acids, amino acids, flavonoids, benzoic acid derivatives, triglycerides, and lipids, showed the main effects of MeJA involved in the significant metabolite pathways. Our study elucidated the changes in rice bean composition and metabolites associated with various stages of sprouting and the roles of MeJA in sprout physiology. Our results can help improve this economically important grain crop.

Background: Xinbao Pill (Xin-Bao-Wan, XBW) is a traditional Chinese herbal formula, which is widely used in clinical treatment for cardiovascular diseases; however, the therapeutic effect of XBW on myocardial ischemia-reperfusion injury (MI/RI) is unclear. In this study, we evaluated the cardioprotective effect and molecular mechanism of XBW against MI/RI.Methods: A phytochemistry-based network pharmacology analysis was used to uncover the mechanism of XBW against MI/RI. Firstly, ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) method was used to identify chemicals in XBW. MI/RI-related targets of XBW were predicted using TargetNet database, OMIC database and reported literatures. GO and KEGG pathway enrichment analyses were performed using String database. Left anterior descending artery (LAD) ligation-induced MI/RI rat model and oxygen glucose deprivation-reperfusion (OGD/R)-induced H9c2 cell model were used to evaluate the cardioprotective effect and potential mechanism of XBW.Results: 37 chemicals were identified in XBW by using UHPLC-QTOF-MS; 50 MI/RI-related targets of XBW were predicted using TargetNet database, OMIC database and reported literatures. In vivo study, XBW (180 mg/kg) significantly reduced myocardial infarct size and creatine kinase MB (CK-MB) level induced by MI/RI in rat model; in vitro study, XBW protected H9c2 cells against OGD/R injury in a dose-dependent manner. GO and KEGG pathway enrichment analyses showed that the cardioprotective effect of XBW was associated with 5 significant pathways including autophagy, apoptosis, HIF-1 signaling pathway, PI3K-Akt signaling pathway, FoxO signaling pathway. Experimental investigation also verified that XBW could suppress apoptosis, autophagy and endoplasmic reticulum (ER) stress.Conclusion: XBW showed therapeutic effects against MI/RI mainly via attenuating apoptosis though suppressing excessive autophagy and ER stress.

Analysis of new psychoactive substances (NPS) still poses a challenge for many institutions due to the number of available substances and the constantly changing drug market. Both new and well-known substances keep appearing and disappearing on the market, making it hard to adapt analytical methods in a timely manner. In this study we developed a qualitative screening approach for serum samples by means of liquid chromatography--quadrupole time-of-flight mass spectrometry. Samples were measured in data-dependent auto tandem mass spectrometry mode and identified by fragment spectra comparison, retention time and accurate mass. Approximately 500 NPS, including 195 synthetic cannabinoids, 180 stimulants, 86 hallucinogens, 26 benzodiazepines and 7 others were investigated. Serum samples were fortified to 1 ng/mL and 10 ng/mL concentrations to estimate approximate limits of identification (LOIs). Samples were extracted using solid-phase extraction with non-endcapped C18 material and elution in two consecutive steps. Benzodiazepines were eluted in the first step, while substances of other NPS subclasses were distributed among both extracts. To determine LOIs, both extracts were combined. Ninety-six percent (470/492) of investigated NPS were detected in 10 ng/mL samples and 88% (432/492) were detected in 1 ng/mL samples. Stimulants stood out with higher LOIs, possibly due to instability of certain methcathinone derivatives. However, considering relevant blood concentrations, the method provided sufficient sensitivity for stimulants as well as other NPS subclasses. Data-dependent acquisition was proven to provide high sensitivity and reliability when combined with an information-dependent preferred list, without losing its untargeted operation principle. Summarizing, the developed method fulfilled its purpose as a sensitive untargeted screening for serum samples and allows uncomplicated expansion of the spectral library to include thousands of targets.

Analysis of synthetic cannabinoids still poses a challenge for many institutions due to the number of available substances and the constantly changing drug market. Both new and well-known substances keep appearing and disappearing on the market, making it hard to adapt analytical methods in a timely manner. In this study, we developed a qualitative screening approach for synthetic cannabinoids and their metabolites by means of liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). Samples were measured in data-dependent auto-MS/MS mode and identified by fragment spectra, retention time and accurate mass. Two established solid phase extractions were compared using fortified serum and urine samples. Mixes of 199 synthetic cannabinoids and 110 metabolites were used in 1- and 10-ng/ml concentrations. Up to 93% of synthetic cannabinoids and 74% of metabolites were detected in fortified 1-ng/ml samples. From February 2018 to October 2020, we analyzed 1492 cases, of which 73 cases were positive for synthetic cannabinoids or metabolites. 5F-MDMB-PICA, 4F-MDMB-BINACA, MDMB-4en-PINACA, and 4F-MDMB-BICA were most frequently detected. Hydrolysis metabolites were detected in many blood samples, providing a longer detection window. Quantification was conducted via liquid chromatography triple quadrupole mass spectrometry after liquid-liquid extraction. Concentrations were mostly close to 1 ng/ml in blood samples. LC-QTOF-MS was able to detect substances above trace quantities (< 0.1 ng/ml) in most cases, therefore fulfilling its purpose as a sensitive general screening approach. Expansion of the screening library was uncomplicated and enables future additions for up to thousands of targets.

Deer antler velvet (DAV) has been traditionally used in Chinese medicine, including treatment on toothache [1] . Due to its rapid and regenerative capacity, deer antlers were proposed to be the good model for bone remodelling in mammals [2] . The data presented in this work is on the liquid chromatography and mass spectrometry (LC-MS) profile and bioactive potential of Malayan deer antler velvet (DAV) on different Candida species that has clinical importance. Aqueous extraction of DAV samples was subjected to Liquid chromatography quadrupole time of flight mass spectrometry (LC-QTOF-MS) profiling. Reverse phase (RP) separation was used due to the process extraction using water as a solvent to separate polar compound. The data was interpreted using Profile Analysis 2.1V. The DAV samples were also tested for the effect on the biofilm formation of seven Candida species in a 96 well plate [3] . The biofilms were developed for 72 h in aerobic environment. Following that, the biofilms biomass was determined using crystal violet assay.

An exhaustive migration study of eight corks, made of ethylene–vinyl acetate, was carried out to identify any non-volatile and volatile compounds using an untargeted approach. The challenge associated with the structural elucidation of unknowns was undertaken using both ultra-high-performance liquid chromatography coupled to an ion-mobility separation quadrupole-time of flight mass spectrometer and gas chromatography mass spectrometry. A total of fifty compounds were observed to migrate from the corks, and among these additives such as antioxidants (Butyl 4-hydroxybenzoate, Irganox 1010, Irganox 1075, Irgafos 168 and BHT) or lubricants (EBO and octadecanamide, N,N'-1,2-ethanediylbis-) were identified. A high proportion (84%) of the detected compounds was non-intentionally added substances (NIAS), and included several cyclic oligomers with different chain sequences. NIAS, such as 2,6-bis(1,1-dimethylethyl)-4-ethyl and 7,9-ditert-butyl-1-oxaspiro[4.5]deca-6,9-diene-2,8-dione, break-down products, including hexa-, hepta- and nonadecanamide, N,N'-1,2-ethanediylbis-, and oxidation products were also identified. One cork was found to be unsuitable for use as a food contact material.

Intact glycopeptide identification has long been known as a key and challenging barrier to the comprehensive and accurate understanding the role of glycosylation in an organism. Intact glycopeptide analysis is a blossoming field that has received increasing attention in recent years. MS-based strategies and relative software tools are major drivers that have greatly facilitated the analysis of intact glycopeptides, particularly intact N-glycopeptides. This article provides a systematic review of the intact glycopeptide-identification process using MS data generated in shotgun proteomic experiments, which typically focus on N-glycopeptide analysis. Particular attention is paid to the software tools that have been recently developed in the last decade for the interpretation and quality control of glycopeptide spectra acquired using different MS strategies. The review also provides information about the characteristics and applications of these software tools, discusses their advantages and disadvantages, and concludes with a discussion of outstanding tools., Graphical abstract, Highlights • Summarize the development of software tools for intact glycopeptide analysis. • Survey the MS acquisition strategies for glycopeptide spectrum acquisition. • Conclude the quality control methods for glycopeptide-spectrum matching. • Show the application of intact glycopeptide analysis in glycoproteomic research., In Brief This article provides a systematic review of the most recent MS-based strategies and corresponding software tools for the analysis of intact glycopeptides, particularly intact N-glycopeptides, reported in the last decade, including the process of identifying N-glycopeptides from MS data, the existing methods of MS data acquisition and interpretation, the quality control methods, the display of results, and the software applications.

A novel and rapid approach to characterise the occurrence of contaminants of emerging concern (CECs) in river water is presented using multi-residue targeted analysis and machine learning-assisted in silico suspect screening of passive sampler extracts. Passive samplers (Chemcatcher®) configured with hydrophilic-lipophilic balance (HLB) sorbents were deployed in the Central London region of the tidal River Thames (UK) catchment in winter and summer campaigns in 2018 and 2019. Extracts were analysed by; (a) a rapid 5.5-min direct injection targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for 164 CECs and (b) a full-scan LC coupled to quadrupole time of flight mass spectrometry (QTOF-MS) method using data-independent acquisition over 15 min. From targeted analysis of grab water samples, a total of 33 pharmaceuticals, illicit drugs, drug metabolites, personal care products and pesticides (including several EU Watch-List chemicals) were identified, and mean concentrations determined at 40 ± 37 ng/L-1. For targeted analysis of passive sampler extracts, 65 unique compounds were detected with differences observed between summer and winter campaigns. For suspect screening, 59 additional compounds were shortlisted based on mass spectral database matching, followed by machine learning-assisted retention time prediction. Many of these included additional pharmaceuticals and pesticides, but also new metabolites and industrial chemicals. The novelty in this approach lies in the convenience of using passive samplers together with machine learning-assisted chemical analysis methods for rapid, time-integrated catchment monitoring of CECs.

Background Reports from experienced heroin users about an alternative and appreciated but harmful so-called “Turkish” heroin preparation technic led to the chemical investigation of the compounds produced during this process and investigation of the presence of other psychoactive contaminants. Methods Comparison of diacetylmorphine, 6-monoacetylmorphine, morphine, paracetamol and caffeine concentrations were performed in the non-processed material, after processing according to the standard and to the alternative preparation methods using liquid chromatography coupled to quadrupole time of flight mass spectrometry followed by statistical evaluation of the results. Results The two preparation methods had in common a diminution of diacetylmorphine as compared to the starting material but significantly more 6-monoacetylmorphine was produced using the “Turkish” preparation method as compared to the standard method. Conclusion The high amount of psychoactive 6-monoacetylmorphine may have an impact on the reported effects of heroin using the “Turkish” preparation procedure.

Objectives Dried ginger and ginger are the same type of medicine and food. The differential components of ginger and dried ginger, dried ginger and ginger charcoal were investigated. Materials and Methods The experimental materials were divided into three sample groups: the ginger group, dried ginger group, and ginger charcoal group. The ginger group, dried ginger group, and ginger charcoal group were qualitatively analyzed by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. The data were processed by Marker View Software. Principal component analysis and orthogonal partial least-square discriminant analysis were performed with SIMCA 13.0 Software. The differential components of the ginger and dried ginger groups as well as the dried ginger and ginger charcoal groups with a variable importance in the projection > 2 (P < 0.05) were identified with PeakView 1.2 Software. Results Ten differential components, including 6-gingerol, 8-gingerol, and 10-gingerol, were identified between the ginger group and dried ginger group; 13 differential components, including 6-shogaol, 10-gingerol, and zingiberone, were identified between the dried ginger group and ginger charcoal group. Conclusions The main differential components between the ginger and dried ginger groups and the dried ginger and ginger charcoal groups were gingerols and diphenylheptanes. Based on metabolomics analysis of the chemical composition of ginger’s medicinal materials, effects, and other related factors, it is recommended that 6-gingerol, 6-shogaol, and zingiberone should be used as indicative components for the respective quality evaluation of ginger, dried ginger and ginger charcoal. The results of this study may provide a basis for the reasonable quality evaluation of ginger medicinal materials.

In this study, the hyphenation of gas chromatography to atmospheric pressure chemical ionization-quadrupole-time-of-flight-mass spectrometry (GC-APCI-Q-TOF-MS) is applied for the investigation of degradation products of ionic liquid (IL) based electrolytes. The advantage of APCI compared to electron ionization (EI) for amine-based analytes due to a higher sensitivity of the molecular ion was demonstrated and the results obtained in this work contribute to a better understanding of the IL aging behavior in regard to their application as green electrolyte for lithium metal batteries (LMBs). Pristine N-butyl-N-methylpyrrolidinium bis(trifluoromethanesulfonyl)imide (Pyr14TFSI) and Pyr14TFSI-based electrolytes with lithium bis(trifluoromethanesulfonyl)imide (LiTFSI) as conducting Li salt were investigated. For this purpose, ion source optimization was performed for amine-based analytes using N-butyl-N-methylbutan-1-amine (BMBA) as standard compound. Furthermore, a customized water flow was directed into the ion source to inhibit in-source reactions, such as fragmentation or oxidation processes, and therefore to promote the [M+H]+-ion formation. The respective headspace (HS) above the ionic liquids (ILs) and electrolytes was sampled at first for the detection of highly volatile analytes. Structure proposals were provided by matching mass spectra obtained from GC-APCI-Q-TOF-MS/MS and GC-EI-MS measurements. Aliphatic amine-, pyrrolidine- and pyrrole-based aging products were identified as decomposition species of the Pyr14 +-cation, e.g. N-butylpyrrole and N-butyl-N-methylpent-4-en-1-amine. Furthermore, the presence of lithium metal on a copper substrate in the pristine IL led to significantly stronger aging effects. Galvanic corrosion processes at the redox-couple Li and Cu were suggested as possible causes. This phenomenon questions the practicability of using copper current collectors with lithium anodes for IL-based battery cell systems. Additionally, the LiTFSI concentration in the electrolyte showed an impact on aging caused by corrosion.

A untargeted metabolomics approach was proposed in this study based on ultra-high performance liquid chromatography quadrupole time-of-flight (UHPLC-QTOF) and rapid evaporative ionization mass spectrometry (REIMS) to discriminate lamb and mutton meat and investigate their subtle metabolic differences, considering the higher popularity of lamb meat than mutton in the market. Multivariate statistical analysis was performed for data processing in order to distinguish between the two sample types. A total of 42 potential metabolites (20 in positive and 22 in negative ion mode) were defined for UHPLC-QTOF analysis, which provided references for discriminating the two kinds of meat. Furthermore, three potential markers were tentatively identified using LC/MS data against chemical databases. In addition, 14 potential metabolites were putatively identified in negative ion mode using the LipidMaps database. Meanwhile, the data-driven soft independent modeling of class analogy (DD-SIMCA) model was established, which could rapidly differentiate non-pretreated lamb meat and mutton with 92% specificity, rendering REIMS a promising technique for meat identification.

Herbal liqueurs are a large group of diverse alcoholic beverages with an ancient tradition produced by maceration of various herbs and spices and are commonly drunken before or after a meal to aid in the digestion because of their potential functional properties. In the presented work, eight different commercial herbal liqueurs were investigated with regard to their composition of phenolic compounds by liquid chromatography ion mobility quadrupole time-of-flight mass spectrometry (LC-IM-QTOF-MS). This multidimensional analytical platform uses all-ion fragmentation for a deep coverage of the foodome. After an extensive data clean-up, 3225 features were found. 213 features were manually annotated due to the absence of databases and software tools able to consider the drift time or Collison Cross Section (CCS) together with high resolution MS/MS spectra for identification. The identified compounds reflected the large variance between the investigated samples and a wealth of potential bioactive compounds that these liqueurs harbor.

The triacylglycerol (TAG) compositions of cucumber, tomato, pumpkin, and carrot seed oils were analyzed using ultra-performance convergence chromatography (UPC2) combined with quadrupole time-of-flight mass spectrometry (Q-TOF MS). A total of 36, 42, 39, and 27 different TAGs were characterized based on their Q-TOF MS accurate molecular weight and MS2 fragment ion profiles in the cucumber, tomato, pumpkin, and carrot seed oils, respectively. Generally, different vegetable seed oils had different TAGs compositions. Among the identified fatty acids, linoleic acid was the most abundant fatty acid in cucumber, tomato, and pumpkin seed oils and the second most abundant in carrot seed oil with relative concentrations of 54.48, 48.69, 45.10, and 15.92 g/100 g total fatty acids, respectively. Oleic acid has the highest concentration in carrot seed oil and the second highest in cucumber, tomato, and pumpkin seed oils, with relative concentrations of 78.97, 18.57, 27.16, and 33.39 g/100 g total fatty acids, respectively. The chemical compositions of TAGs and fatty acids could promote understanding about the chemical profiles of certain vegetable seed oils, thus improving the potential ability to select appropriate oils with specific functions and a high nutritional value and then develop functional foods in the future.

Triterpene saponins (TSs) are important bioactive constituents with structural diversity widely distributed in many plants. The root of Phytolacca acinosa Roxb (RPa) has been used as a traditional Chinese medicine. However, TSs as the main active ingredients in RPa have not been fully characterized. Here, we profiled TSs from RPa by high-performance liquid chromatography coupled to electrospray ionization and quadrupole time-of-flight mass spectrometry (HPLC-ESI-QTOF-MS/MS). We tentatively identified 29 TSs, including 13 that had not been reported previously from this plant. This study indicates that HPLC-ESI-QTOF-MS/MS is an effective and rapid method for the characterization of complicated TSs in herbal extracts.

Mycotoxin contamination is an increasing concern in aquafeed industry due to the replacement of marine-based ingredients to plant-based ingredients and climate changes associated with growing mycotoxin contamination in plant ingredients. Consequently, the health and growth of aqua animals could be affected by contaminated fish feeds, and finally influence human health. Therefore, it is necessary to develop an effective screening method for mycotoxin determination in fish feed. This master project develops two screening methods (using low resolution UHPLC-QqQ-MS/MS and high resolution UHPLC-IMS-QTOF MS instruments) for the determination of 18 mycotoxins: aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2), beauvericin (BEA), deoxynivalenol (DON), diacetoxyscirpenol (DAS), enniatin A (ENA), enniatin A1 (ENA1), enniatin B (ENB), enniatin B1 (ENB1), fumonisin B1 (FB1), fumonisin B3 (FB3), moniliformin (MON), ochratoxin A (OTA), T2-Toxin (T2), HT-2 Toxin (HT2) and neosolaniol (NEO) and 6 metabolites: 15-Acetyldeoxynivalenol (15-ADON), 3-Acetyldeoxynivalenol (3-ADON), alpha-zearalenol (αZEL), beta-zearalenol (βZEL), deoxynivalenol-3-glucoside (D3G) and deepoxy deoxynivalenol (DOM-1). In addition to screening method, a quantitative method was developed by UHPLC-QqQ-MS/MS for all compounds except AFs, MON and DAS. The developed method was validated in terms of linearity. ENA, ENA1, ENB, ENB1, BEA, NEO, HT-2, T-2 and αZEL had a R2 > 0.99; DON, 3-ADON, OTA and βZE had a R2 >0.95, showing good linearity performance. Recovery (95%-111%) and intra-day precision (RSD of 4%-18%) for ENA, ENA1, ENB, ENB1 and BEA using T-2 as IS, demonstrated excellent validation performances. Both isotopic IS and structural analogue IS were used during the validation. T-2 was confirmed as a good alternation to substitute expensive isotopic IS for the validation of ENA, ENA1, ENB, ENB1 and BEA. Ten fish feeds including one fish feed ingredient collected in the market were analyzed using the screening method. Results show considerable mycotoxin contamination for all samples. The developed method by UHPLC-QqQ has shown great promise in quantification of mycotoxins. The other method was developed by UHPLC-IMS-QTOF MS for all compounds with CCS value as extra identification point. The developed method was applied to ten commercial fish feeds including one fish feed ingredient and results reveals considerable mycotoxin contamination for almost all samples. Mycotoxins detected in the fish feed samples were ENNs (including ENA, ENA1, ENB and ENB1) and BEA, while FUMs (FB1 and FB3) were detected in the fish feed ingredient sample. A chemical degradation experiment for two frequently occurred mycotoxins (BEA and ENB) was also performed. The transformation products (TPs) of both compounds are tentatively predicted. Master's Thesis in Chemistry KJEM399 MAMN-KJEM

Breath analysis is a promising method for metabolomics studies and clinical diagnosis, as it enables the observation of metabolites in a convenient and noninvasive way. In this work, an atmospheric pressure photoionization (APPI) source was modified for online analysis of exhaled breath by coupling with quadrupole time-of-flight mass spectrometry (QTOFMS). Three parameters, namely, the capillary voltage, the sampling flow and the curtain gas flow of the APPI source, were optimized. Five healthy volunteers, three males and two females, were enrolled to test the performance of modified APPI-QTOFMS by analyzing their exhaled breath. A total of 21 compounds were tentatively identified, and four metabolites, namely, dimethyl selenoxide, δ-valerolactam, hydroxymandelic acid and palmitic amide were detected in the exhaled breath for the first time. The result shows that modified APPI-QTOFMS can be used for the online study of exhaled breath. Graphical abstract.

Helichrysum arenarium (L.) Moench, a traditional herbal medicine distributed in Europe and Central Asia, has been used to lower blood lipids and treat obesity. However, the active compounds of H. arenarium, and the potential mechanism by which they reduce obesity remain unclear. The primary objective of this study was to identify the anti-obesity compounds in H. arenarium extracts. Based on the compound library of Helichrysum species, 55 compounds, including six with anti-obesity activity, have been identified by ultrafiltration affinity and ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UF-UPLC-Q-TOF-MS). Interestingly, all of the detected active compounds were derivatives of caffeoylquinic acid. Among them, isochlorogenic acid B showed the most potent inhibitory activity toward lipase, with an IC50 value of 12.63 ± 2.99 μg/mL. The potential mechanisms of the six active compounds for the treatment of obesity were explored. This work presents an efficient method for rapid screening lipase inhibitors from complex natural products, which can be easily adapted to other therapeutically relevant enzymatic systems. Additionally, the results revealed the chemical basis and mechanism of the anti-obesity properties of H. arenarium.

The aroma fraction of fresh fruits is a key factor to evaluate quality and acceptance or resistance of customers. Due to the limited sensitivity of current analytical methods, many important aroma components are unidentified in most studies of fresh fruit. In this work, the aroma compounds in the fresh navel orange were studied by multidimensional gas chromatography quadrupole time-of-flight mass spectrometry which can simultaneously achieve one-dimensional and comprehensive two-dimensional separation on only one instrument, with no need to change columns. As a result, a total of 97 volatile compounds were determined by comprehensive two-dimensional gas chromatography while only 44 compounds were identified by one-dimensional gas chromatography. The identified compounds, which account for 98.30% of the volatile chemicals present in the orange, include 10 aldehydes, 8 ketones, 22 alcohols, 12 esters, 11 hydrocarbons and 34 terpenes. D-limonene was shown to be the predominant compound in fresh navel oranges. The results show that comprehensive two-dimensional gas chromatography provided reliable and comprehensive information on the aroma compounds in fresh navel oranges.

We developed a metabolomics workflow using ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry to determine the effect of thermal treatment on milk composition and metabolites based on multivariate data analysis. We analyzed raw, pasteurized, and UHT milk samples. The samples were first centrifuged to remove the fat layer and mixed with methanol to precipitate proteins. Subsequently, the supernatant was analyzed by ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry in electrospray negative mode. Mass spectral data were acquired in MSE mode, a technique whereby both precursor and fragment mass spectral are simultaneously acquired by alternating between low and high collision energy (CE) during a single analytical run, to enable metabolite identification. Based on multivariate data analysis, these markers were significantly affected by thermal treatment. Among the 8 potential markers, we identified 7 oxylipids (9-hydroxydecanoic acid, 12-hydroxydodecanoic acid, 2-hydroxymyristic acid, 3-hydroxytetradecanoic acid, 5-hydroxyeicosatetraenoic acid, 3-hydroxyhexadecanoic acid, and 10-hydroxyoctadecanoic acid) and 1 phospholipid (LysoPE, hexadecanoyl-lysophosphatidylethanolamine). The oxylipids seemed to be adequate for distinguishing UHT milk from raw and pasteurized milk. The structures of the 8 potential markers were identified and characterized using informatics software. Our metabolomics workflow provides a fast approach for the identification of various types of milk.

The tentative identification of bioactive compounds in the extract of Vernonia amygdalina leaf was carried out using positive ionization of Liquid chromatography-mass spectrometry quadrupole time of flight (LC-Q-TOF/MS). The positive ionization is associated with the presence of saponins, flavonoids, alkaloids, terpenoids, and glycosides. Tentative assignments of the secondary metabolites were performed by comparing the MS fragmentation patterns with Waters® UNIFY library which allows positive identification of the compounds based on the spectral match. All the metabolites compounds were estimated and presented in a BPI (Base peak intensity) plot. These data are the unpublished supplementary materials related to “Ethanolic extraction of bioactive compounds from V. amygdalina leaf using response surface methodology as an optimization tool” (Alara et al., 2018).

Ciguatera Fish Poisoning (CFP) is primarily caused by consumption of tropical and sub-tropical fish contaminated by Ciguatoxins (CTXs). These lipid-soluble, polyether neurotoxins are produced by dinoflagellates in the genera Gambierdiscus and Fukuyoa. While there is no regulatory level in Europe for CTXs, the European Food Safety Authority (EFSA) adopted the United States guidance level of 0.01 μg P-CTX1B eq.kg−1 of fish. This limit is extremely low and requires significant improvement in the detection of CTXs. In this study, we compared analytical protocols based on liquid chromatography coupled to tandem low or high resolution mass spectrometry (LC-LRMS or HRMS) to find the best conditions for sensitivity and/or selectivity. Different approaches such as LC conditions, ion choice and acquisition modes, were evaluated to detect the Pacific-ciguatoxins (P-CTXs) on a triple quadrupole (API4000 Qtrap, Sciex) or a quadrupole time of flight (QTOF 6550, Agilent Technologies) spectrometer. Moreover, matrix effects were calculated using matrix-matched calibration solutions of P-CTX1B and P-CTX3C prepared in purified fish extract. Subsequently, the method performance was assessed on naturally contaminated samples of seafood and phytoplankton. With LRMS, the ammoniated adduct ion used as a precursor ion showed an advantage for selectivity through confirmatory transitions, without affecting signal-to-noise ratios, and hence limits of detection (LODs). As also reported by some studies in the literature, methanol-based mobile phase gave better selectivity and sensitivity for the detection of P-CTXs. While the LOD for P-CTX1B and P-CTX3C met the EFSA recommendation level when using LRMS, the findings suggested careful evaluation of instrumental parameters for determination of CTXs. LODs were significantly higher for HRMS, which currently results in the need for a significantly higher sample intake. Nevertheless, HRMS allowed for the identification of artefacts and may allow for improved confirmation of the identity of P-CTXs analogues. Consequently, LRMS and HRMS are considered complementary to ensure adequate quantitation and identification of P-CTXs.

Using high-performance liquid chromatography coupled to high-resolution electrospray ionization quadrupole time-of-flight mass spectrometry (HPLC/ESI-Q-TOF-MS), we have developed a new method for detection and identification of furan fatty acids (F-acids), which are widely distributed in living organisms and foods as minor lipid components and are known to have antioxidant and anti-inflammatory effects. For this purpose, total fatty acids prepared from the testis lipids of Japanese chum salmon (Oncorhynchus keta) were examined without any concentration or isolation of F-acids. In negative ESI mode, F-acids gave a prominent [M−H]− ion, by which individual F-acids could be detected and identified. High-resolution extracted ion chromatograms clearly showed the occurrence of five major F-acid homologs as already reported by GC/MS. The method was successfully applied to several fish samples and revealed the occurrence of F-acids for the first time in the two New Zealand fish, hoki (Macruronus novaezelandiae) and school shark (Galeorhinus galeus).

Comprehensive screening procedures for psychoactive agents in body fluids are an essential task in clinical and forensic toxicology. With the continuous emergence and adaption of new psychoactive substances (NPS) keeping a screening method up to date is challenging. To meet these demands, hyphenated high-resolution mass spectrometry has gained interest as extensive and expandable screening approach. Here we present a comprehensive method for systematic toxicological analysis of serum by liquid chromatography-quadrupole-time-of-flight mass spectrometry (LC-QTOF-MS) with data independent acquisition. The potential of this method was demonstrated by analysis of 247 authentic serum- and 12 post-mortem femoral blood samples. Thus 950 compounds, comprising 185 different drugs and metabolites could be identified. For the detected substances, including pharmaceutical substances, illicit drugs as well as NPS, serum concentrations were confirmed ranging from traces to toxic values indicating the capability for forensic toxicological requirements. Positive identification of drugs was achieved by accurate mass measurement (±5 ppm for [M + H]+; ±10 ppm for [M − H]−), retention time (±0.35 min), isotopic pattern match (less than 10 m/z RMS [ppm]), isotope match intensity (less than 20% RMS) and the presence of at least two fragment ions. The LC-QTOF-MS procedure was shown to be superior to serum screening by GC–MS, since 240% (335 versus 141) more drugs were identified in serum samples compared to GC–MS.