Fluid Chromatography Tandem Mass Spectrometry Research Articles

Analysis of unsaturated fatty acids by supercritical fluid chromatography tandem mass spectrometry coupled with online Paternò-Büchi reaction

Carbon double bond (CC) positions in unsaturated fatty acids (UFAs) can be identified based on Paternò-Büchi (PB) reaction coupled with mass spectrometry (MS). However, achieving a high conversion rate and confusing mass spectrum are still challenging. In this study, an online PB reaction coupled with supercritical fluid chromatography-mass spectrometry (SFC-MS), using a homemade flow microreactor, was established to analyze CC bonds in UFAs, and benzaldehyde (BA) was proposed as the optimized reagent. Benefiting from this setup, both prototype and product ions of FAs can be rapidly captured at the same retention time, significantly enhancing the accuracy of lipid identification, especially when dealing with complex compounds. Furthermore, a high CC bond conversion rate was achieved, ranging from 53 to 67 %. Additionally, this strategy demonstrated excellent relative quantification abilities, which were based on the ratios of either the sum of diagnostic ions or the individual diagnostic ions. This novel approach was proposed and applied for the specific recognition of free FA in human plasma, which successfully provides quantitative information about UFA isomers. This strategy is believed to hold promise for the analysis of unsaturated lipid isomers in complex biological samples.

Rapid enantiomeric analysis of zopiclone in serum by supercritical fluid chromatography-tandem mass spectrometry.

Zopiclone (ZOP) is a hypnotic drug prescribed to treat insomnia. Due to the chiral nature of ZOP, the psychologically active S-form and inactive R-form need to be determined enantiomerically in a forensic drug analysis. In the present study, a supercritical fluid chromatography (SFC) method was designed with a faster analysis ability than that of previously reported techniques. The SFC-tandem mass spectrometry (SFC-MS/MS) method was optimized using a column with a chiral polysaccharide stationary phase (Trefoil CEL2). ZOP was extracted from pooled human serum using solid-phase extraction (Oasis HLB) and analyzed. The developed SFC-MS/MS method achieved the baseline separation of S-ZOP and R-ZOP within 2 min. The fit-for-purpose method validation indicated that the optimized solid-phase extraction achieved near complete recovery and approximately 70% of the matrix effect. Both the retention time and peak area showed sufficient precision. The lower and upper limits of quantification (LOQ) were 5.7 × 10-2  ng/mL and 25 ng/mL for R-ZOP, and 5.2 × 10-2  ng/mL and 25 ng/mL for S-ZOP. The calibration line was linear in the range from lower LOQ to upper LOQ. The stability test indicated that ZOP in serum stored in a refrigerator (4°C) degraded and about 55% remained in 31 days. The quick analysis of the SFC-MS/MS method makes it a valid option for the enantiomeric analysis of ZOP.

Determination of amphetamine enantiomers in urine by conductive vial electromembrane extraction and ultra-high performance supercritical fluid chromatography tandem mass spectrometry.

Separation and quantification of amphetamine enantiomers are commonly used to distinguish between consumption of prescription amphetamine (mostly S-amphetamine) and illicit forms of the drug (racemate). In this study, electromembrane extraction with prototype conductive vials was combined with ultra-high performance supercritical fluid chromatography (UHPSFC-MS/MS) to quantify R- and S-amphetamine in urine. Amphetamine was extracted from 100 μL urine, diluted with 25 μL internal standard solution and 175 μL 130 mM formic acid, across a supported liquid membrane (SLM) consisting of 9μL of a 1:1(w/w) mixture of 2-nitrophenyloctyl ether (NPOE) and bis(2-ethylhexyl)phosphite (DEHPi) into an acceptor phase containing 300 μL 130 mM formic acid. The extraction was facilitated by the application of 30 V for 15 min. Enantiomeric separation was achieved using UHPSFC-MS/MS with a chiral stationary phase. The calibration range was 50-10,000 ng/mL for each enantiomer. The between-assay CV was ≤5%, within-assay CV ≤ 1.5%, and bias within ±2%. Recoveries were 83%-90% (CV ≤ 6%), and internal standard corrected matrix effects were 99-105 (CV ≤ 2%). The matrix effects ranged from 96% to 98% (CV ≤ 8%) when not corrected by the internal standard. The EME method was compared with a chiral routine method that employed liquid-liquid extraction (LLE) for sample preparation. Assay results were in agreement with the routine method, and the mean deviation between methods was 3%, ranging from -21% to 31%. Finally, sample preparation greenness was assessed using the AGREEprep tool, which resulted in a greenness score of 0.54 for conductive vial EME, opposed to 0.47 for semi-automated 96-well LLE.

Levels and distribution profiles of Per- and Polyfluoroalkyl Substances (PFAS) in a high Arctic Svalbard ice core

Per- and polyfluoroalkyl substances (PFAS) are a group of persistent organic contaminants of which some are toxic and bioaccumulative. Several PFAS can be formed from the atmospheric degradation of precursors such as fluorotelomer alcohols (FTOHs) as well as hydrochlorofluorocarbons (HFCs) and other ozone-depleting chlorofluorocarbon (CFC) replacement compounds. Svalbard ice cores have been shown to provide a valuable record of long-range atmospheric transport of contaminants to the Arctic. This study uses a 12.3 m ice core from the remote Lomonosovfonna ice cap on Svalbard to understand the atmospheric deposition of PFAS in the Arctic. A total of 45 PFAS were targeted, of which 26 were detected, using supercritical fluid chromatography (SFC) tandem mass spectrometry (MS/MS) and ultra-performance liquid chromatography (UPLC) MS/MS. C2 to C11 perfluoroalkyl carboxylic acids (PFCAs) were detected continuously in the ice core and their fluxes ranged from 2.5 to 8200 ng m−2 yr−1 (9.51–16,500 pg L−1). Trifluoroacetic acid (TFA) represented 71 % of the total mass of C2 – C11 PFCAs in the ice core and had increasing temporal trends in deposition. The distribution profile of PFCAs suggested that FTOHs were likely the atmospheric precursor to C8 – C11 PFCAs, whereas C2 – C6 PFCAs had alternative sources, such as HFCs and other CFC replacement compounds. Perfluorooctanesulfonic acid (PFOS) was also widely detected in 82 % of ice core subsections, and its isomer profile (81 % linear) indicated an electrochemical fluorination manufacturing source. Comparisons of PFAS concentrations with a marine aerosol proxy showed that marine aerosols were insignificant for the deposition of PFAS on Lomonosovfonna. Comparisons with a melt proxy showed that TFA and PFOS were mobile during meltwater percolation. This indicates that seasonal snowmelt and runoff from post-industrial accumulation on glaciers could be a significant seasonal source of PFAS to ecosystems in Arctic fjords.

Development and application of a SFC–DAD–MS/MS method to determine carotenoids and vitamin A in egg yolks from laying hens supplemented with β-carotene

β-Carotene, a provitamin A carotenoid, can be converted into vitamin A in animals’ bodies, and can also be accumulated intactly in many animal products. In this study, supercritical fluid chromatography–tandem mass spectrometry was utilized to determine β-carotene and different forms of vitamin A in eggs simultaneously. According to the results, β-carotene contained in yolk reached a plateau after about 2 weeks of supplementation. With an increase in dietary supplement level, the amount of β-carotene gradually increased, as well as slightly changing the yolk color. Moreover, the contents of retinoids including retinol, retinyl propionate, retinyl palmitate and retinyl stearate were also elevated in yolks with the β-carotene additive levels; meanwhile, the lutein and zeaxanthin decreased. On the whole, β-carotene in the diet of laying hens could be partially deposited in egg yolk, and the contents of vitamin A in yolk could be increased due to β-carotene bioconversion.

Comprehensive evaluation of novel fungicide benzovindiflupyr at the enantiomeric level: Bioactivity, toxicity, mechanism, and dissipation behavior

Racemates in the environment can lead to inaccurate risk assessment. To obtain the enantiomeric level information of benzovindiflupyr for accurate risk assessment, the absolute configuration of benzovindiflupyr was first confirmed, and the enantioseparation method was developed by supercritical fluid chromatography tandem mass spectrometry. The enantioselectivity for bioactivity and toxicity was investigated, and the mechanism was explored by molecular docking and detecting succinate dehydrogenase (SDH) activity and content of succinate acid. 1S,4R-(−)-benzovindiflupyr was identified as the most active against the six targeted phytopathogens, which showed higher 1.7–54.5 times than 1R,4S-(+)-benzovindiflupyr. Additionally, 1S,4R-(−)-benzovindiflupyr (LD50: 21.54 μg L−1) was 103.7 times more toxic than 1R,4S-(+)-benzovindiflupyr against Daphnia magna. 1S,4R-(−)-benzovindiflupyr had a stronger affinity for SDH and significantly inhibited SDH activity, resulting in an increase in succinate acid in the tricarboxylic acid cycle, while its downstream products, fumaric and L-malic acid were significantly reduced. Moreover, the dissipation behavior of benzovindiflupyr on three vegetables was evaluated. 1S,4R-(−)-benzovindiflupyr was preferentially degraded in tomato, but opposite in leaves. The enantioselectivity in pepper and cucumber leaves was the same as in tomato, while there was no enantioselectivity in pepper and cucumber. The study provides a basis for accurate risk assessment and the development of high-effective and low-risk fungicides.

Fast and Ultrasensitive Glycoform Analysis by Supercritical Fluid Chromatography-Tandem Mass Spectrometry.

The glycoform of a therapeutic monoclonal antibody (mAb) has a significant impact on its effector function as well as its safety and pharmacokinetics. Glycoform heterogeneity is influenced by various factors, including the producing cells and cell culture processes. Therefore, accurate glycoform characterization is essential for drug design, process optimization, manufacturing, and quality control of therapeutic mAbs. In this study, we developed a fast, quantitative, and highly sensitive analytical platform for glycan profiling by supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS) and applied the technique to the glycan structural analysis of mAbs. To achieve both the highest sensitivity and the most comprehensive glycan profiling, we integrated our energy-resolved oxonium ion monitoring (Erexim) method with SFC-MS to construct a new SFC-Erexim technology. An 8 min analysis of bevacizumab, nivolumab, ramucirumab, rituximab, and trastuzumab by SFC-Erexim detected a total of 102 glycoforms, with a detection limit of 5 attomoles. The dynamic range of glycan abundance was over 6 orders of magnitude for bevacizumab analysis by SFC-Erexim compared to 3 orders of magnitude for conventional fluorescence HPLC analysis. This method revealed the glycan profile characteristics and lot-to-lot heterogeneity of various therapeutic mAbs. We were also able to detect a series of structural variations in pharmacologically important glycan structures. The SFC-MS-based glycoform profiling method will provide an ideal platform for the in-depth analysis of precise glycan structure and abundance.

Development of a Chiral Supercritical Fluid Chromatography-Tandem Mass Spectrometry and Reversed-Phase Liquid Chromatography-Tandem Mass Spectrometry Platform for the Quantitative Metabolic Profiling of Octadecanoid Oxylipins.

Octadecanoids are broadly defined as oxylipins (i.e., lipid mediators) derived from 18-carbon fatty acids. In contrast to the well-studied eicosanoids, there is a lack of analytical methods for octadecanoids, hampering further investigations in the field. We developed an integrated workflow combining chiral separation by supercritical fluid chromatography (SFC) and reversed-phase liquid chromatography (LC) coupled to tandem mass spectrometry detection for quantification of a broad panel of octadecanoids. The platform includes 70 custom-synthesized analytical and internal standards to extend the coverage of the octadecanoid synthetic pathways. A total of 103 octadecanoids could be separated by chiral SFC and complex enantioseparations could be performed in <13 min, while the achiral LC method separated 67 octadecanoids in 13.5 min. The LC method provided a robust complementary approach with greater sensitivity relative to the SFC method. Both methods were validated in solvent and surrogate matrix in terms of linearity, lower limits of quantification (LLOQ), recovery, accuracy, precision, and matrix effects. Instrumental linearity was good for both methods (R2 > 0.995) and LLOQ ranged from 0.03 to 6.00 ng/mL for SFC and 0.01 to 1.25 ng/mL for LC. The average accuracy in the solvent and surrogate matrix ranged from 89 to 109% in SFC and from 106 to 220% in LC, whereas coefficients of variation (CV) were <14% (at medium and high concentrations) and 26% (at low concentrations). Validation in the surrogate matrix showed negligible matrix effects (<16% for all analytes), and average recoveries ranged from 71 to 83%. The combined methods provide a platform to investigate the biological activity of octadecanoids and expand our understanding of these little-studied compounds.

Optimization and validation of a fast supercritical fluid chromatography tandem mass spectrometry method for the quantitative determination of a large set of PFASs in food matrices and human milk

An Ultra-High Performance Supercritical Fluid Chromatography coupled with tandem Mass Spectrometry analytical method (UHPSFC-MS/MS) was developed for the determination of 34 perfluoroalkylated substances (PFASs) in food-related matrices. Two parameters (i.e. stationary phase and co-solvent) were selected and optimized using a step-by-step method, while a design of experiment (DoE) method using a central composite design (CCD) was implemented to optimize column temperature, mobile phase flow rate, co-solvent concentration and automated back pressure regulator (ABPR). The Torus 2-PIC column was selected along with ammonium acetate AcoNH4 as additive in the co-solvent. DoE optimization of both peak width and resolution enabled validating an optimized model (desirability 0.613) and setting column temperature at 38.7 °C, AcoNH4 concentration at 8 mM, mobile phase flow rate of 1.9 mL/min and ABPR at 1654 psi. The validated resulting method enabled reaching limits of quantification below 0.2 ng/g (w.w.) for 97 % PFASs in accordance with current EU requirements. The strategy was successfully applied to the characterization of a range (n > 30) of food-related matrices (red meat, poultry meat, eggs, fish and breast milk) collected in Algeria in 2019. PFOA and PFBA were observed as the most frequently detected PFASs, i.e. in 96.96 % and 90.9 % of the samples respectively. The highest concentrations were determined in fishery products up to 4.42 ng/g (w.w.) for PFTeDA and 0.75 ng/g (w.w.) for PFOS.

Rapid Simultaneous Quantification of 1-Formyl-2,2-Dimethylhydrazine and Dimethylurea Isomers in Environmental Samples by Supercritical Fluid Chromatography–Tandem Mass Spectrometry

When released to the environment, the rocket fuel unsymmetrical dimethylhydrazine (UDMH) undergoes oxidative transformations, resulting in the formation of an extremely large number of nitrogen-containing transformation products, including isomeric compounds which are difficult to discriminate by common chromatography techniques. In the present work, supercritical fluid chromatography–tandem mass spectrometry (SFC-MS/MS) was proposed for resolving the problem of fast separation and simultaneous quantification of 1-formyl-2,2-dimethylhydrazine (FADMH) as one of the major UDMH transformation products, and its isomers—1,1-dimethylurea (UDMU) and 1,2-dimethylurea (SDMU). 2-Ethylpyridine stationary phase provided baseline separation of analytes in 1.5 min without the distortion of the chromatographic peaks. Optimization of SFC separation and MS/MS detection conditions allowed for the development of rapid, sensitive, and “green” method for the simultaneous determination of FADMH, UDMU, and SDMU in environmental samples with LOQs of 1–10 µg L−1 and linear range covering three orders of magnitude. The method was validated and successfully tested on the real extracts of peaty and sandy soils polluted with rocket fuel and UDMH oxidation products. It was shown that both UDMU and SDMU are formed in noticeable amounts during UDMH oxidation. Despite relatively low toxicity, UDMU can be considered one of the major UDMH transformation products and a potential marker of soil pollution with toxic rocket fuel.

Simultaneous fatal poisoning of two victims with 4F-MDMB-BINACA and ethanol

PurposeMethyl-2-(1-(4-fluorobutyl)-1H-indazole-3-carboxamido)-3,3-dimethylbutanoate (4F-MDMB-BINACA) is a newly emerging synthetic cannabinoid receptor agonists (SCRA) first described in 2018 in both Europe and the United States. Two fatal cases are reported caused by simultaneous consumption of 4F-MDMB-BINACA and ethanol.MethodsThe victims were brothers who were both found deceased after consuming 4F-MDMB-BINACA and ethanol. Post-mortem toxicological analyses of blood and urine were carried out by supercritical fluid chromatography tandem mass spectrometry (SFC–MS/MS) and headspace gas chromatography with flame ionization detection (HS-GC–FID).ResultsThe concentration of 4F-MDMB-BINACA in the postmortem blood was 2.50 and 2.34 ng/mL, and blood alcohol concentration was 2.11 and 2.49 g/L, respectively.ConclusionAccording to the reported cases and reviews of the scientific literature, concurrent ethanol consumption should amplify the toxicity of SCRAs. The threshold SCRA concentration for fatal overdose can be estimated ng/mL level (0.37–4.1 ng/mL according to the reported cases) in cases in which 1.5–2.5 g/L of ethanol is present in the blood.

Separation and determination of fluindapyr enantiomers in cucumber and tomato and by supercritical fluid chromatography tandem mass spectrometry

Fluindapyr is a pyrazolamide chiral fungicide of succinate dehydrogenase inhibitor (SDHIs) with two enantiomers. Pesticide enantiomers often exhibit different biological activities, toxicity due to their different enantioselectivity. Therefore, it is important to separate fluindapyr enantiomers and assess each enantiomer. In this study, fluindapyr enantiomers were baseline separated by supercritical fluid chromatography-mass spectrometry in 2 min. The limit of quantification (LOQ) of this method was 5 μg/kg. The developed method was applied to monitor the fluindapyr enantiomers in cucumber and tomato, the data showed that R-(-)-fluindapyr was preferentially degraded in tomato leaves, S-(+)-fluindapyr was preferentially degraded in cucumber leaves, and fluindapyr enantiomers had no enantioselective degradation behavior in two fruits. It is proved again that enantiomers have different enantioselective degradation behavior with the different plant species and even to different parts of the same plant. The enantioselectivity is likely to be caused by different biodegradation enzyme systems.

Comparative Metabolomics of Small Molecules Specifically Expressed in the Dorsal or Ventral Marginal Zones in Vertebrate Gastrula.

Many previous studies have reported the various proteins specifically secreted as inducers in the dorsal or ventral regions in vertebrate gastrula. However, little is known about the effect on cell fate of small molecules below 1000 Da. We therefore tried to identify small molecules specifically expressed in the dorsal marginal zone (DMZ) or ventral marginal zone (VMZ) in vertebrate gastrula. Small intracellular and secreted molecules were detected using explants and supernatant samples. Hydrophilic metabolites were analyzed by capillary ion chromatography–mass spectrometry and liquid chromatography–mass spectrometry, and lipids were analyzed by supercritical fluid chromatography–tandem mass spectrometry. In total, 190 hydrophilic metabolites and 396 lipids were identified. The DMZ was found to have high amounts of glycolysis- and glutathione metabolism-related metabolites in explants, and the VMZ was richer in purine metabolism-related metabolites. We also discovered some hydrophilic metabolites and lipids differentially contained in the DMZ or VMZ. Our research would contribute to a deeper understanding of the cellular physiology that regulates early embryogenesis.

Supercritical Fluid Chromatography-Tandem Mass Spectrometry for Rapid Quantification of Pentacyclic Triterpenoids in Plant Extracts.

Pentacyclic triterpenoids (PCTs) are a widely distributed class of plant secondary metabolites. These compounds have high bioactive properties, primarily antitumor and antioxidant activity. In this study, a method was developed for the quantitative analysis of pentacyclic triterpenoids in plants using supercritical fluid chromatography–tandem mass spectrometry (SFC-MS/MS). Separation of ten major PCTs (friedelin, lupeol, β-amyrin, α-amyrin, betulin, erythrodiol, uvaol, betulinic, oleanolic and ursolic acids) was studied on six silica-based reversed stationary phases. The best results (7 min analysis time in isocratic elution mode) were achieved on an HSS C18 SB stationary phase using carbon dioxide—isopropanol (8%) mobile phase providing decisive contribution of polar interactions to the retention of analytes. It was shown that the use of atmospheric pressure chemical ionization (APCI) is preferred over atmospheric pressure photoionization (APPI). The combination of SFC with APCI-MS/MS mass spectrometry made it possible to achieve the limits of quantification in plant extracts in the range of 2.3–20 μg·L−1. The developed method was validated and tested in the analyses of birch outer layer (Betula pendula) bark, and licorice (Glycyrrhiza glabra) root, as well as lingonberry (Vaccinium vitis-idaea), cranberry (Vaccinium oxycoccos), apple (Malus domestica “Golden Delicious” and Malus domestica “Red Delicious”) peels.

Enantioseparation and dissipation of acephate and its highly toxic metabolite methamidophos in pakchoi by supercritical fluid chromatography tandem mass spectrometry.

Acephate is widely used in crops as racemate. However, the enantioselective dissipation of acephate enantiomers has not been investigated in pakchoi. A sensitive and effective approach was established for determining residues of acephate and its highly toxic metabolite methamidophos enantiomers by supercritical fluid chromatography tandem mass spectrometry. Baseline separations for their enantiomers were achieved by using a Chiralcel OD-H column. The optimal chromatographic conditions were obtained as follows: CO2 /ethanol (95/5) as mobile phase; flow rate, 3.0mL/min; column temperature, 40°C. The mean recoveries (RSDs) of analytes were in the range of 77-83.1% (6.1-9.9%), 75.4-87.5% (9.3-13.2%), and 81.5-84.2% (7.1-13.4%) at three fortification levels (0.005, 0.05, and 0.5mg/kg for each enantiomer) for interday assay (n=18). The method was used to evaluate the enantioselective dissipation of acephate and methamidophos in pakchoi. S-acephate dissipated faster than R-acephate, while the concentration of R-methamidophos was higher than that of S-methamidophos during the entire study period. The results indicated that the R-enantiomer of acephate and methamidophos was preferentially enriched in pakchoi. The established analysis approach and the study data provided useful information for the rational use of acephate in agriculture.

Simultaneous Determination of Vitamin D and Its Hydroxylated and Esterified Metabolites by Ultrahigh-Performance Supercritical Fluid Chromatography–Tandem Mass Spectrometry

In this study, an analytical method has been developed that, for the first time, allows simultaneous determination of vitamin D2 and vitamin D3 along with their hydroxylated and esterified forms. A group of 12 vitamin D analogues including vitamin D2 and vitamin D3, seven hydroxylated metabolites, and three ester forms were separated in a single 8.0 min run using ultrahigh-performance supercritical fluid chromatography coupled with triple quadrupole tandem mass spectrometry. Electrospray ionization and atmospheric pressure chemical ionization were investigated as ion sources, of which the latter showed a higher ionization efficiency. Chromatographic conditions were thoroughly evaluated by a step-by-step method, whereas an experimental design was applied for the optimization of the ionization parameters. Calibration and repeatability studies were carried out to validate the instrumental methodology showing determination coefficients higher than 0.9992 and good intra- and interday precision with relative standard deviations for areas and retention times lower than 10 and 2.1%, respectively, for all target analytes. Limits of quantification were below 3.03 μg/L for all compounds. The methodology was then validated and applied for the evaluation of human plasma samples in order to demonstrate its applicability to the analysis of vitamin D analogues in biological samples. Samples of five individuals were analyzed. Results show that linoleate-D3, vitamin D2, vitamin D3, 25-hydroxyvitamin D2, 24,25-dihydroxyvitamin D3, and 1,25-dihydroxyvitamin D3 could be detected in most samples, while the two latter also were quantified in all analyzed samples.

Evaluation of matrix effect in determination of mevinphos stereoisomers in tobacco by supercritical fluid chromatography tandem mass spectrometry

Supercritical fluid chromatography tandem mass spectrometry (SFC-MS/MS) is an important tool for the analysis of stereoisomer in complex matrices. Matrix effect is a critical factor that can significantly compromise the accuracy of pesticide residue analyses by mass spectrometry. The influence of matrix effect in the stereoselective analysis of mevinphos in tobacco by SFC-MS/MS was studied. Experiments have been performed to compare the response of mevinphos isomers in standard solutions prepared in acetonitrile with their responses in tobacco extracts. The presence and extent of matrix effects are simultaneously affected by several factors (matrix concentration, matrix type, matrix dilution, and chromatographic separation conditions). In this study, a QuEChERS method coupled with a dilution strategy eliminated most of the matrix effects. The mean recoveries for all isomers were 85.2%–99.3%, with 2.2%–7.7% intra-day relative standard deviations (RSDs) and 3.6%–6.9% inter-day RSDs at 0.05, 0.2 and 1.0 mg/kg levels. Good coefficients of determination (R2 ≥ 0.9995) were achieved for all studied isomers over the concentration range of 25–500 ng/mL.

Supercritical fluid chromatography tandem mass spectrometry employed with evaporation-free liquid-liquid extraction for the rapid analysis of cinnarizine in rat plasma.

Cinnarizine is a weak base, which can produce supersaturation and precipitation during gastrointestinal transit, affecting its absorption in vivo. Therefore, it is necessary to investigate whether the oral bioavailability of cinnarizine can be improved after co-administration with precipitation inhibitors or not. In order to evaluate the pharmacokinetic behavior of cinnarizine in rats, a simple, rapid, sensitive, and environmentally friendly supercritical fluid chromatography-tandem mass spectrometric method was established and validated. In this method, flunarizine, a structural analogue of cinnarizine, was selected as the internal standard, and cinnarizine was extracted from rat plasma using evaporation-free liquid-liquid extraction method. The analytes were separated on a Torus 1-AA column (3.0mm × 100mm, 1.7 μm) within 2.0min, using a gradient elution procedure. The transitions of cinnarizine and flunarizine were m/z 369.1 → 167.1 and m/z 405.1 → 203.1, respectively. Cinnarizine showed good linear correlation in the range of 1-500ng/ml with a lower limit of quantification of 1ng/ml. The intra- and interday precision and accuracy of all quality control samples were within ±15%. This high-throughput, accurate, sensitive, and reproducible method has been successfully applied to study the effects of the precipitation inhibitor cinnarizine on the pharmacokinetics in rats.

Enantioselective monitoring chiral fungicide mefentrifluconazole in tomato, cucumber, pepper and its pickled products by supercritical fluid chromatography tandem mass spectrometry

A fast, effective, and environmental-friendly method was developed for enantioseparation and analysis of mefentrifluconazole in vegetables based on supercritical fluid chromatography tandem mass spectrometry. The enantioselective behaviors of mefentrifluconazole enantiomers in tomato, cucumber, and pepper in the greenhouse, and pickled cucumber and pepper during processing were investigated. Mefentrifluconazole enantiomers could obtain baseline separation within 2 min. The average recoveries of all matrices ranged from 78.4% to 119.0%, with relative standard deviations less than 16.8% for two enantiomers. S-(+)-mefentrifluconazole was preferentially degraded in pepper, while there was no enantioselectivity in tomato and cucumber under field conditions. During processing, S-(+)-mefentrifluconazole was reduced preferentially than R-(−)-mefentrifluconazole in pickled cucumber and cucumber brine. Inversely, R-(−)-mefentrifluconazole degraded faster than S-(+)-mefentrifluconazole in pepper brine. But, no obvious enantioselectivity was observed in pickled pepper. The result of this study could contribute to a more accurate dietary risk assessment of mefentrifluconazole in vegetables and processed products.

Comparison of Kit-Based Metabolomics with Other Methodologies in a Large Cohort, towards Establishing Reference Values.

Metabolic profiling is an omics approach that can be used to observe phenotypic changes, making it particularly attractive for biomarker discovery. Although several candidate metabolites biomarkers for disease expression have been identified in recent clinical studies, the reference values of healthy subjects have not been established. In particular, the accuracy of concentrations measured by mass spectrometry (MS) is unclear. Therefore, comprehensive metabolic profiling in large-scale cohorts by MS to create a database with reference ranges is essential for evaluating the quality of the discovered biomarkers. In this study, we tested 8700 plasma samples by commercial kit-based metabolomics and separated them into two groups of 6159 and 2541 analyses based on the different ultra-high-performance tandem mass spectrometry (UHPLC-MS/MS) systems. We evaluated the quality of the quantified values of the detected metabolites from the reference materials in the group of 2541 compared with the quantified values from other platforms, such as nuclear magnetic resonance (NMR), supercritical fluid chromatography tandem mass spectrometry (SFC-MS/MS) and UHPLC-Fourier transform mass spectrometry (FTMS). The values of the amino acids were highly correlated with the NMR results, and lipid species such as phosphatidylcholines and ceramides showed good correlation, while the values of triglycerides and cholesterol esters correlated less to the lipidomics analyses performed using SFC-MS/MS and UHPLC-FTMS. The evaluation of the quantified values by MS-based techniques is essential for metabolic profiling in a large-scale cohort.