Gas Chromatography-mass Spectrometry Detection Research Articles

Comparative study of chemical constituents and biological potency in Lantana camara Linn. leaf essential oil collected from two different sites in Northeast India

The ubiquitous occurrence of Lantana camara highlights its exceptional ecological versatility. Leaves of L. camara were obtained for this study from two different locations of NE, India. Hydro distillation was used to extract the essential oils from the leaves, and gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detection (GC-FID) were used to study the chemical profiles of the isolated oils. The essential oil analysis detected 35 components in total essential oil from Location 1 (BLCEO) while in Location 2 essential oil (JLCEO), 29 components were identified. Caryophyllene, humulene and sabinene were present in considerably major amounts in both BLCEO and JLCEO, though variation in area percentage was seen. The anti-oxidant properties of BLCEO were better than that of JLCEO. The BLCEO has also exhibited better results in inhibiting acetylcholinesterase. However, both the essential oils did not show any prominent results in α-amylase inhibitory assay and anti-tyrosinase activity. In this study, the phytochemical composition and their effect on the bioactivity of both samples were observed. The chemical constitution of L. camara essential oils (LCEO) and its possible ecological ramifications have also been well explained.

Investigating the effects of microplastics on the metabolism of Trematomus bernacchii from the ross sea (Antarctica)

Microplastic pollution is a growing environmental issue, even reaching remote areas like the Arctic and Antarctic, posing threats to biodiversity and food chains. The present research represents a pioneering endeavor aimed at exploring the relationship between lipids and microplastics in 20 wild specimens of Trematomus bernacchii from the Ross Sea (Antarctica). Fish were grouped in MPs-ingested and MPs-free based on whether they had ingested microplastics. Raman spectroscopy revealed that contaminated fish samples contained from one to three different types of polymeric fibers (1.4 items/specimen, ± = 0.7), specifically, polyester (PES), polypropylene (PP), and polyethylene terephthalate (PET). Gas chromatography-mass spectrometry (GC–MS) and gas chromatography-flame ionization detection (GC-FID) techniques were employed for the study of the lipid composition in term of fatty acids methyl esters (FAMEs). Fifty different FAME compounds were identified and quantified in the lipid fraction extracted from the muscle tissues of the selected fish samples. Polyunsaturated fatty acids (PUFAs) are the most abundant family of fatty acids in T. bernacchii species with eicosapentaenoic (C20:5ω3) and docosahexaenoic (C22:6ω3) acids as main components. In detail, PUFA class accounted for 46.78 ± 6.82% and 44.62 ± 4.86% of the total fatty acid composition in MPs-ingested and MPs-free groups, respectively. The contents of the monounsaturated fatty acids (MUFAs) and saturated fatty acids (SFAs) varied from 27.93% to 31.15% and from 24.23% to 25.05% in MPs-ingested and MPs-free fish samples, respectively. Based on Mann-Whitney test results (p < 0.05), there was no significant difference from a statistical point of view between two groups of fishes. Additionally, nutritional quality indices exhibited comparable values between groups. Results showed that no significant differences were found in the fatty acids distribution between the two groups. This indicates that the lipid composition of wild fish that are naturally exposed to plastic pollution remains unchanged and could still have beneficial effects on human health.

Artificial intelligence tools and concepts give access to authenticity and quality information in Brazilian olive oil volatilome

Brazil is the world's second largest importer of olive oil and stared to produce its own oil quite recently in two distinct geographic areas. The production is still very small, and studies regarding oil composition are scarce. Determination of discriminant differences are useful to support appeals regarding origin indication and to detect fraud and adulteration with oils from other countries. In this work the Brazilian extra virgin olive oils (EVOOs) volatilome is explored by Artificial Intelligence (AI) tools developed on multiple headspace solid-phase microextraction (MHS-SPME) combined with comprehensive 2D gas chromatography-mass spectrometry and flame ionization detection (GC×GC-MS/FID) data. Using MHS-SPME, external standard calibration, and FID predicted relative response factors (RRF), the accurate quantification of 51 informative volatile compounds was carried out and the odorants responsible for key-positive sensory attributes were used to generate distinctive aroma blueprints aligned with the AI smelling based on sensomics. As authenticity and origin assessment decision-making tool, augmented visualization by computer vision was applied on volatilome 2D fingerprints. By this approach, extra-virgin olive oil samples (n=35), from two olive cultivars (Arbequina and Koroneiki) harvested in the main producing regions in Brazil (Rio Grande do Sul and Serra da Mantiqueira) in 2021 and 2022, were effectively discriminated and mislabeled products regarding their geographical origin were, for the first time, promptly identified.

Monitoring studies on contamination of urban runoff with hazardous benzotriazoles and bisphenols in one of the least polluted places worldwide

Benzotriazoles (BTR) and bisphenols (BP) are artificial contaminants of emerging concern (CECs) commonly found in everyday products. This study focuses on urban runoff to investigate the occurrence of BTRs and BPs in Iceland, regarded as one of the least polluted places in the world, which made it reasonable to confirm or deny the presence of these micropollutants in its environment. Samples collected in February 2023 (SC1) and August 2024 (SC2) from seven locations along Iceland's Ring Road were evaluated to determine the occurrence of seven BTRs (1H-BTR, 4Me-BTR, 5Me-BTR, 5Cl-BTR, UV-P, UV-326, UV-329) and six BPs (BPF, BPE, BPA, BPZ, BPAP, BPM) in the runoff, using the ultrasound-assisted emulsification-microextraction for analytes isolation and gas chromatography-mass spectrometry for detection (USAEME-GC/MS). All locations showed detectable and varying levels of BTRs and BPs, with 5Cl-BTR (11.6 μg/L) and BPF (56.3 μg/L), both in SC1, demonstrating the highest concentrations, providing valuable insights into their prevalence and distribution. A correlational analysis investigated the connection between these pollutants and various characteristics associated with the locations along the Ring Road. This study contributes to the essential comprehension of these CECs, serving as input for future strategies for monitoring and mitigating their impact.

Determination of multiple restricted and prohibited ingredients in hair dyes using a novel 3D stationary phase coupled with gas chromatography-mass spectrometry

Hair dyes (HDs) are mainly composed of various benzene series, amines, and phenolic compounds. These ingredients are well known to have allergenic, teratogenic, and carcinogenic properties. As such, the presence of these ingredients in HDs has received increased attention in recent years. At present, the applications of traditional analytical and detection methods and commercial chromatographic columns are limited by problems such as poor qualitative analysis and inaccurate quantification. Thus, the development of new analytical and detection technologies and stationary phases is an urgent endeavor. Moreover, HDs contain complex compounds and exhibit significant matrix interference. Hence, appropriate sample pretreatment methods are necessary to analyze HDs. In this study, the 3D nonpolar rigid structure of triptycene (TP) was combined with the polar flexible chains of polyethylene glycol (PEG) to design and synthesize a TP derivative, TP-PEG, as a stationary phase for chromatographic columns. The stationary phase enabled the expansion of the selection range for polar and nonpolar analytes. Subsequently, gas chromatography-mass spectrometry (GC-MS) was used to quantitatively analyze 22 ingredients in HDs. The experimental results demonstrated that analytes with different polarities exhibited sharp and symmetrical peak shapes on the stationary phase, and all 22 analytes achieved baseline separation on the chromatographic column. The 22 ingredients in HDs showed good linear relationships within their respective ranges, with correlation coefficients greater than 0.9985. The average recovery rates at three spiked levels were in the range of 89.2%-103.2%, and RSDs were less than 5%. Compared with traditional methods, the proposed method has higher efficiency and better accuracy, thus verifying the excellent separation performance of the new stationary phase and the effectiveness of the established GC-MS detection method. The findings indicated the applicability of the developed method to the detection and analysis of various compounds in HDs.

Isolation and characterization of Thymus serpyllum L. essential oil and mass fragmentation analysis of major constituents

ABSTRACT The essential oils were isolated from Thymus serpyllum L. leaves, stem, flowers and whole plant using hydrodistillation technique and % yield was determined. The chemical constituents were studied using Gas Chromatography-Mass Spectrometry and Gas Chromatography-Flame Ionisation Detection. The analysis revealed the presence of 38 compounds in the essential oils obtained from different plant parts. Identification of compounds was achieved by detailed analysis of mass spectrum and retention indices. The major constituents of essential oil from the whole plant were thymol (35.25%), p-cymene (27.57%), thymol acetate (8.03%) and γ-terpinene (6.92%). In leaves they were thymol (45.59%), p-cymene (24.26%), thymol acetate (6.41%), and β-bisabolene (5.91%); whereas in flowers these were thymol (68.74%) and thymol acetate (7.40%) and in the stem, these were thymol (55.15%) and thymol acetate (8.01%). Specifically, it is the first report wherein essential oil constituents of T. serpyllum L. whole plant, leaves, flowers, and stems have been analyzed separately. Further, fundamental analysis of complex mass fragmentation patterns of terpenes and terpenoids using EI-MS has been achieved and the logical reason for the fragmentation pattern has been elaborated. Detailed elaboration of the mass fragmentation pattern may strengthen the EI-MS-based compound identification of terpenes and terpenoids from essential oils.

Simultaneous determination of sixteen phthalic acid esters (PAEs) in soil and evaluation of matrix effect using a QuEChERS/GC/MS-internal standard method.

Phthalic acid esters (PAEs) are emerging pollutants that need to be analyzed precisely. Chromatography-based determination of PAE content in soils are frequently affected by matrix effect, which may limit the quantification of different kinds of PAEs from different types of soil. Here we optimized a QuEChERS protocol combined with gas chromatography-mass spectrometry (GC-MS) for simultaneous determination of 16 PAEs in different soils. PAEs in different type of soils (fluvo-aquic soil, red soil, and black soil) were extracted with acetonitrile followed by GC-MS detection based on quantitative ion internal standard method. All 16 PAEs showed excellent linear relationships with mass peak areas (R2 > 0.99). The limits of detection (LOD) and limits of quantitation (LOQ) of all the samples were in the range of 0.91-66.97µg/kg and 2.7-200.9µg/kg, respectively. The accurate test at 0.5, 0.1, and 1.0mg/kg spiking level recorded recovery rate between 80.11% and 100.99% with relative standard deviations (RSDs) ranging from 0.37 to 8.50% in tested matrices. No significant matrix effect was observed for most tested PAEs. This is a simple method with high sensitivity and strong stability, which is suitable and reproducible for quantifying large number of PAEs in different types of soil.

Chemical components and antioxidant potential of essential oil of Rhanterium epapposum growing in sandy habitat in Saudi Arabia: In silico study on cytochrome P450 1A1 and xanthine oxidase

Rhanterium epapposum Oliv. (Family: Asteraceae) is a common plant in Saudi Arabia, possessing a variety of traditional uses. The current targets were studying the comparative chemical profile and antioxidant of the essential oil (EO) of R. epapposum collected from a sandy habitat at Qareenah, Riyadh, Saudi Arabia. The EO profiling was constructed via gas chromatography-mass spectrometry (GC-MS) and GC-flame ionization detection (GC-FID). Forty-eight compounds were found, including monoterpenes (hydrocarbons 3.25% and oxygenated 48.64%), sesquiterpenes (hydrocarbons 12.64 % and oxygenated 32.38%), which are the backbone of the oil along with a minor of other non-terpenoid compounds (1.31%). Linalool (18.02%), α-eudesmol (11.98%), trans-sesquisabinene hydrate (11.63%), aromadendrene oxide-(1) (7.68%), nerol (6.88%), and α-terpineol (5.01%) were identified as the dominant components. The EO was evaluated for its antioxidant activity using in vitro DPPH, ABTS, and FRAP methods. The results demonstrated that R. epapposum EO possesses a dosage-dependent ability for antioxidant scavenging activity as per all three techniques. This EO exhibited IC50 values of 517.59, 350.24, and 713.72 μg/mL for the DPPH, ABTS, and FRAP, respectively, through the comparison with the ascorbic acid with respective values of 36.10, 26.12, and 44.58 μg/mL. The main compounds were subjected to docking analysis on cytochrome P450 1A1 and xanthine oxidase. All evaluated compounds exhibited good binding affinities especially α-eudesmol and linalool for both enzymes, respectively. The current research confirmed the potent free radical scavenging properties of R. epapposum EO. More research on the antioxidant content of this oil and/or its primary constituents in pure form on the two enzymes, cytochrome P450 1A1 and xanthine oxidase, is proposed to develop a better understanding of the oxidative action routes in vivo and in vitro.

Baltic Sea shipwrecks as a source of hazardous pollution

Background Shipwrecks on the Baltic Sea seabed pose a serious threat to the marine environment. Fuel, ammunition and chemicals in their holds can enter the ecosystem at any time, causing an ecological disaster. It is known that oil spills from ship accidents can affect life and health of different species of animals, both immediately after catastrophe and for many years thereafter. This article discusses the negative impact of shipwrecks on the ecological status of the Baltic Sea and presents the contamination status of bottom sediment core samples taken in the vicinity of shipwrecks located in the South Baltic, i.e., S/s Stuttgart, t/s Franken, S/T Burgmeister Petersen and m/s Sleipner. It is based on the results of research carried out by the Maritime Institute between 2011 and 2016. Methods Core samples were taken by a VKG-2 vibrating probe and analysed towards content of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), total petroleum hydrocarbons (TPHs) and total organic carbon (TOC). Seven PAHs and Seven PCBs were determined using solid phase extraction (SPE) technique followed by gas chromatography-mass spectrometry (GC-MS) detection. In order to determine the concentration of TPH, the SPE technique was applied followed by gas chromatography with flame ionisation detector (GC-FID) analysis. TOC content was established by TOC analyser using infrared detection. Results Samples taken in the vicinity of the S/s Stuttgart shipwreck have shown the highest concentrations of all analysed parameters compared to the examined wrecks and significantly differed from the results typical for these areas. This studies have shown that the S/s Stuttgart shipwreck poses a serious threat to the marine environment of Baltic Sea and it is necessary to continue studies in this area and to perform a wider range of analysis.

Unravelling the correlations among the microbial community, physicochemical properties, and volatile compounds of traditional mung bean sour liquid

Sour liquid noodles are a characteristic traditional food in Luoyang, China. Mung bean sour liquid (MBSL) is fermented from mung beans and used to cook noodles (instead of using water). However, the microbial community, volatile compounds, and physicochemical properties of MBSL are still unclear. Here, a comprehensive study was conducted using a combination of Illumina HiSeq sequencing, gas chromatography‒mass spectrometry detection, and physicochemical analysis methods. Eight phyla and 59 genera of bacteria were identified in MBSL, of which 4 genera were dominant, namely, Lactobacillus, Lactococcus, Leuconostoc, and Lelliottia. Six phyla and 123 genera of fungi were detected, with 11 genera being dominant. The protein content, total acid content, and pH mainly influenced the microbial community. Myroides, Aeromonas, Flavobacterium, and Lactobacillus, etc. May play important roles in balancing MBSL ecosystems. There were 40 volatile compounds in MBSL, with 1-hexanol, eugenol, acetic acid, and hexanoic acid being considered key volatile compounds. Lactobacillus, Algoriella, Lelliottia, and Leuconostoc were significantly correlated with the volatile compounds found in MBSL. The complex correlation between microbial communities, volatile compounds, and physicochemical properties may be elucidated through these findings. Controlling environmental factors and increasing the number of selected microbial cultures may improve the quality of MBSL.

Electrophysiological and Behavioral Responses of Cabbage Aphid (Brevicoryne brassicae) to Rosemary (Rosmarinus officinalis) Volatiles, a Potential push Plant for Vegetable push-pull Cropping System.

The cabbage aphid (Brevicoryne brassicae) is a major pest of kale (Brassica oleraceae var. acephala), an important vegetable that is grown worldwide due to its high nutritional and economic value. Brevicoryne brassicae poses a great challenge to B. oleraceae var. acephala production, causing significant direct and indirect yield losses. Farmers overly rely on synthetic insecticides to manage the pest with limited success owing to its high reproductive behavior and development of resistance. This necessitates a search for sustainable alternatives to mitigate these challenges. This study assessed behavioral responses of B. brassicae to odors from rosemary (Rosmarinus officinalis) and B. oleraceae var. acephala headspace volatiles in a Perspex four-arm olfactometer. We identified and quantified volatiles emitted by each of the two plants and those eliciting antennal response using coupled gas chromatography-mass spectrometry (GC-MS) and GC-electroantennograhic detection(GC-EAD), respectively. Our findings revealed that B. brassicae spent more time in the arms of the olfactometer that contained B. oleraceae var. acephala volatiles compared to the arm that held R. officinalis volatiles. Additionally, B. brassicae spent more time in the olfactometer arms with B. oleracea var. acephala compared to the arms holding B. oleracea var. acephala and R. officinalis enclosed together and clean air. GC-MS analysis revealed diverse and higher quantities of volatile compounds in R. officinalis compared to B. oleraceae var. acephala. GC-EAD analysis showed that antennae of B. brassicae detected Linalool, α-Terpineol, Verbenone, Geraniol, Camphor, and Borneol from the volatiles of R. officinalis, and Sabinene, γ-Terpinene, and β-Caryophyllene from B. oleraceae var. acephala volatiles.Our findings demonstrate the potential of R. officinalis as a repellent plant against B. brassicae and could be utilized as a 'push' plant in an intercropping strategy against this pest.

Changes in GC-MS metabolite profile, antioxidant capacity and anthocyanins content during fermentation of fine-flavor cacao beans from Ecuador.

The fermentation of fine-flavor cacao beans is a key process contributing to the enhancement of organoleptic attributes and monetary benefits for cacao farmers. This work aimed to describe the dynamics of the gas chromatography-mass spectrometry (GC-MS) metabolite profile as well as the antioxidant capacity and anthocyanin contents during fermentation of fine-flavor cacao beans. Samples of Nacional x Trinitario cacao beans were obtained after 0, 24, 48, 72, 96, and 120 hours of spontaneous fermentation. Total phenolic content (TPC), ferric reducing antioxidant power (FRAP), and total anthocyanin content were measured by ultraviolet-visible (UV-Vis) spectrophotometry. Volatiles were adsorbed by headspace solid phase microextraction (HS-SPME) while other metabolites were assessed by an extraction-derivatization method followed by gas chromatography-mass spectrometry (GC-MS) detection and identification. Thirty-two aroma-active compounds were identified in the samples, including 17 fruity, and 9 floral-like volatiles as well as metabolites with caramel, chocolate, ethereal, nutty, sweet, and woody notes. Principal components analysis and Heatmap-cluster analysis of volatile metabolites grouped samples according to the fermentation time. Additionally, the total anthocyanin content declined during fermentation, and FRAP-TPC values showed a partial correlation. These results highlight the importance of fermentation for the improvement of the fine-flavor characteristics of cacao beans.

Therapeutic effect of Sanhua decoction on rats with middle cerebral artery occlusion and the associated changes in gut microbiota and short-chain fatty acids.

Sanhua decoction (SHD), a traditional prescription, has long been used in treating ischemic stroke (IS). However, the therapeutic effect of SHD and the associated changes in gut microbiota and short-chain fatty acids (SCFAs) are uncertain. In this study, a rat model of IS was established by the middle cerebral artery occlusion (MCAO). By evaluating the cerebral infarct area and brain tissue pathology, it was found that SHD ameliorated IS-related symptoms in MCAO rats. Using 16S rRNA gene sequencing, we found that SHD reduced abnormally elevated Lactobacillus and opportunistic pathogens such as Desulfovibrio, but increased some beneficial bacteria that produce SCFAs, including Clostridia, Lachnospiraceae, Ruminococcaceae, and Coprococcus. KEGG analysis revealed that SHD regulates several pathways, including D-arginine and D-ornithine metabolism, polyketide sugar unit biosynthesis, and cyanoamino acid metabolism, which are significantly altered in MCAO rats. By gas chromatography-mass spectrometry detection of SCFAs, we found that fecal acetic acid, valeric acid, and caproic acid were significantly increased in MCAO rats, whereas propionic acid and isobutyric acid were decreased. SHD reversed the changes in acetic acid and propionic acid in the model rats and significantly increased fecal butyric acid. In addition, MCAO rats had significantly higher serum levels of acetic acid, butyric acid, isovaleric acid, and valeric acid, and lower levels of caproic acid. Altered serum levels of butyric acid, isovaleric acid, valeric acid, and caproic acid were restored, and the level of isobutyric acid was reduced after SHD administration. Spearman analysis revealed that cerebral infarct area had a strong correlation with Bifidobacterium, Desulfovibrio, Lachnospiraceae, Lactobacillus, acetic acid, valeric acid, and caproic acid. Overall, this study demonstrates for the first time that the effect of SHD on IS may be related to gut microbiota and SCFAs, providing a potential scientific explanation for the ameliorative effect of SHD on IS.

Detection of a bibenzyl core scaffold in 28 common mangrove and associate species of the Indian Sundarbans: potential signature molecule for mangrove salinity stress acclimation.

Bibenzyl derivatives comprising two benzene rings are secondary plant metabolites with significant therapeutic value. To date, bibenzyl derivatives in the Plant kingdom have been primarily identified in bryophytes, orchids, and Cannabis sativa. The metabolic cost investment by plant species for the synthesis of these bioactive secondary metabolites is rationalized as a mechanism of plant defense in response to oxidative stress induced by biotic/abiotic factors. Bibenzyl derivatives are synthesized from core phenylpropanoid biosynthetic pathway offshoots in plant species. Mangrove and mangrove associate species thrive under extreme ecological niches such as a hypersaline intertidal environment through unique adaptive and acclimative characteristics, primarily involving osmotic adjustments followed by oxidative stress abatement. Several primary/secondary bioactive metabolites in mangrove species have been identified as components of salinity stress adaptation/acclimation/mitigation; however, the existence of a bibenzyl scaffold in mangrove species functioning in this context remains unknown. We here report the confirmed detection of a core bibenzyl scaffold from extensive gas chromatography-mass spectrometry and gas chromatography-flame ionization detection analyses of 28 mangrove and mangrove associate species from the Indian Sundarbans. We speculate that the common presence of this bibenzyl core molecule in 28 mangrove and associate species may be related to its synthesis via branches of the phenylpropanoid biosynthetic pathway induced under high salinity, which functions to detoxify reactive oxygen species as a protection for the maintenance of plant metabolic processes. This finding reveals a new eco-physiological functional role of bibenzyls in unique mangrove ecosystem.

An accurate and sensitive determination of selected pesticides in mixed fruit juice samples using the combination of a simple and efficient microextraction method and GC-MS with a matrix matching calibration strategy.

Pesticides have been used on several fruits, vegetables and cereals to control harmful organisms in order to increase the quality of products; however, these substances cause serious health effects. Therefore, an accurate and sensitive analytical method should be developed for the determination of pesticides to evaluate their toxicity. In this study, an efficient microextraction strategy was applied to preconcentrate eight different selected pesticides from mixed fruit juice samples prior to gas chromatography-mass spectrometry detection. All significant parameters such as spraying number, extraction solvent type, sample volume and mixing type/period belonging to the developed extraction method were elaborately optimized to get low detection limits. After the optimization studies, system analytical performance studies were carried out and limit of detection (LOD) values varied from 0.04 μg /kg-1 to 1.99 μg kg-1 (mass based) for the selected analytes. Under the optimum experimental conditions, spiking recovery experiments were performed in the mixed fruit juice samples to evaluate the applicability and accuracy of the proposed method. The recovery results were recorded in the range of 81.4-123.5% with acceptable standard deviations by applying a matrix matching calibration strategy. The proposed analytical method can be used for the qualitative and quantitative determination of selected pesticides in the mixed fruit juice samples and can also be applied to other fruit juice samples using a matrix matching calibration strategy.

Identification and validation of diagnostic biomarkers for intrahepatic cholestasis of pregnancy based on untargeted and targeted metabolomics analyses of urine metabolite profiles

BackgroundIntrahepatic cholestasis of pregnancy (ICP) is a prevalent pregnancy-specific complication that presents with maternal itching and elevated serum bile acid levels. ICP is associated with unfavorable pregnancy outcomes, severely decreasing the pregnant woman’s quality of life. Timely identification of ICP is crucial for effective management and improved outcomes.MethodsWe collected urine samples from 8 patients with ICP and 8 healthy individuals. We used Liquid Chromatography-Mass Spectrometry (LC-MS) to detect metabolite expression levels, then conducted a series of bioinformatic analyses to explore the potential biological meanings of differentially expressed metabolites, and preliminarily discovered several candidate biomarkers. To validate these candidate biomarkers, we performed Gas Chromatography-Mass Spectrometry (GC-MS) detection and analyzed their diagnostic values using receiver operating characteristic (ROC) curve.ResultsUntargeted metabolomics data showed that 6129 positive peaks and 6218 negative peaks were extracted from each specimen. OPLS-DA analysis and the heat map for cluster analysis showed satisfactory capability in discriminating ICP specimens from controls. Subsequent analysis extracted 64 significantly differentially expressed metabolites, which could be potential biomarkers for diagnosis of ICP. Based on the KEGG enrichment analyses, six candidate biomarkers were preliminarily identified. Two most promising biomarkers (3-hydroxypropionic acid and uracil) were validated by targeted metabolomics analyses with the area under the curve (AUC) of 0.920 and 0.850 respectively.ConclusionBased on preliminary screening from untargeted metabolomics and subsequent validation through targeted metabolomics, 3-hydroxypropionic acid and uracil were identified as promising diagnostic biomarkers for ICP.

A small-scale silica gel column chromatography method for separating carbazole compounds from highly mature crude oil

The concentration of carbazoles in highly mature crude oil is quite low, making it challenging to separate carbazole compounds for the gas chromatography-mass spectrometry (GC–MS) detection. This study presents a small-scale column chromatography method for separating carbazoles from highly mature crude oil using silica gel as a solid phase adsorbent and a Pasteur pipette as a separation device. The carbazole-rich crude oil from the Pearl River Mouth Basin was selected to explore the impact of reagent polarity and injection mode on the separation of carbazoles. The oil sample was eluted with solvents mixed with different volume proportions of n-hexane and dichloromethane and each eluted fraction was collected for GC–MS testing. The results indicated that increasing the reagent polarity caused the aromatic hydrocarbons and carbazole compounds in crude oil to be eluted sequentially. Most aromatic compounds in the crude oil could be selectively eluted using a reagent polarity ratio of 9:1 (Vn-hexane: Vdichloromethane), with no carbazole compounds. A significant amount of carbazole compounds were eluted in the polar segments of 8:2–6:4, with the eluted carbazoles concentration accounting for more than 98 % of the total concentration. Moreover, the concentration and recovery of carbazoles eluted by direct injection mode were about 10 % higher than those after adsorption by silica gel. The standard deviation of the parameter ratio for the separated carbazole compounds in the three groups of repeatable parallel experiments was less than 0.2 %. Our method is superior to traditional two-step method and C18 column method in separation efficiency and damage to human body. This method can be applied to both highly mature crude oil and other kinds of oils including biodegradable oil. It could be a versatile method for the carbazoles separation and provide technical support in unveiling the geochemical implications of these compounds in complex areas.

Gas Chromatography-Mass Spectrometry Detection of Thymoquinone in Oil and Serum for Clinical Pharmacokinetic Studies.

Thymoquinone (TQ) is the primary component of Nigella sativa L. (NS) oil, which is renowned for its potent hepatoprotective effects attributed to its antioxidant, anti-fibrotic, anti-inflammatory, anti-carcinogenic, and both anti- and pro-apoptotic properties. The aim of this work was to establish a method of measuring TQ in serum in order to investigate the pharmacokinetics of TQ prior to a targeted therapeutic application. In the first step, a gas chromatography-mass spectrometry method for the detection and quantification of TQ in an oily matrix was established and validated according to European Medicines Agency (EMA) criteria. For the assessment of the clinical application, TQ concentrations in 19 oil preparations were determined. Second, two serum samples were spiked with TQ to determine the TQ concentration after deproteinization using toluene. Third, one healthy volunteer ingested 1 g and another one 3 g of a highly concentrated NS oil 30 and 60 min prior to blood sampling for the determination of serum TQ level. After the successful establishment and validation of the measurement method, the highest concentration of TQ (36.56 g/L) was found for a bottled NS oil product (No. 1). Since a capsule is more suitable for oral administration, the product with the third highest TQ concentration (No. 3: 24.39 g/L) was used for all further tests. In the serum samples spiked with TQ, the TQ concentration was reliably detectable in a range between 5 and 10 µg/mL. After oral intake of NS oil (No. 3), however, TQ and/or its derivatives were not detectable in human serum. This discrepancy in detecting TQ after spiking serum or following oral ingestion may be attributed to the instability of TQ in biomatrices as well as its strong protein binding properties. A pharmacokinetics study was therefore not viable. Studies on isotopically labeled TQ in an animal model are necessary to study the pharmacokinetics of TQ using alternative modalities.

Optimization of Dieldrin Selection for the Genetic Sexing of Aedes albopictus.

The mass production of mosquitoes at an industrial scale requires efficient sex separation, which can be achieved through mechanical, genetic or artificial intelligence means. Compared with other methods, the genetic sexing approach offers the advantage of limiting costs and space by removing females at the larval stage. We recently developed a Genetic Sexing Strain (GSS) in Aedes albopictus based on the sex linkage of the rdlR allele, conferring resistance to dieldrin, to the male (M) locus. It has been previously reported that dieldrin ingested by larvae can be detected in adults and bioaccumulated in predators, raising the question of its use at a large scale. In this context, we performed several experiments aiming at optimizing dieldrin selection by decreasing both dieldrin concentration and exposure time while maintaining a stable percentage of contaminating females averaging 1%. We showed that the previously used dieldrin exposure induced an important toxicity as it killed 60% of resistant males at the larval stage. We lowered this toxicity by reducing the dose and/or the exposure time to recover nearly all resistant males. We then quantified the residues of dieldrin in resistant male adults and showed that dieldrin toxicity in larvae was positively correlated with dieldrin concentrations detected in adults. Interestingly, we showed that the use of reduced dieldrin exposure led to a dieldrin quantification in adult males that was below the quantity threshold of the Gas Chromatography-Mass Spectrometry detection method. Presented data show that dieldrin exposure can be adjusted to suppress toxicity in males while achieving efficient sexing and lowering the levels of dieldrin residues in adults to barely quantifiable levels.

Flowering phenophases influence the antibacterial and anti-biofilm effects of Thymus vulgaris L. essential oil

BackgroundEssential oils are becoming increasingly popular in medicinal applications because of their antimicrobial effect. Thymus vulgaris L. (Lamiaceae) is a well-known and widely cultivated medicinal plant, which is used as a remedy for cold, cough and gastrointestinal symptoms. Essential oil content of thyme is responsible for its antimicrobial activity, however, it has been reported that the chemical composition of essential oils influences its biological activity. In order to explore flowering phenophases influence on the chemical composition of thyme essential oil and its antibacterial and anti-biofilm activity, plant materials were collected at the beginning of flowering, in full bloom and at the end of flowering periods in 2019.MethodsEssential oils from fresh and dried plant materials were distilled and analyzed with gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detection (GC-FID). The antibacterial activity was performed by broth microdilution and thin layer chromatography-direct bioautography (TLC-DB) assays and the anti-biofilm effect by crystal violet assay, respectively. Scanning electron microscopy was applied to illustrate the cellular changes of bacterial cells after essential oil treatment.ResultsThymol (52.33–62.46%) was the main component in the thyme essential oils. Thyme oil distilled from fresh plant material and collected at the beginning of flowering period exerted the highest antibacterial and anti-biofilm activity against Haemophilus influenzae, H. parainfluenzae and Pseudomonas aeruginosa.ConclusionThe different flowering periods of Thymus vulgaris influence the antibacterial and anti-biofilm activity of its essential oils, therefore, the collection time has to be taken into consideration and not only the full bloom, but the beginning of flowering period may provide biological active thyme essential oil.