Supercritical Fluid Chromatography-tandem Mass Spectrometry Research Articles

Quantification of Octadecanoids in Human Plasma Using Chiral Supercritical Fluid Chromatography-Tandem Mass Spectrometry.

Octadecanoids are a subset of oxylipins derived from 18-carbon fatty acids. These compounds have historically been understudied but have more recently attracted attention to their purported biological activity. One obstacle to the study of octadecanoids has been a lack of specific analytical methods for their measurement. A particular limitation has been the need for chiral-based methods that enable separation and quantification of individual stereoisomers. The use of chirality provides an additional dimension for distinguishing analytes produced enzymatically from those formed through autoxidation. In this chapter, we describe a comprehensive method using chiral supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS) for the quantification of octadecanoids in human plasma. This method stands as an effective approach for quantifying octadecanoids and is applicable to diverse research applications including clinical research.

Determination of Macrolides in Water Using Solid-Phase Extraction and Supercritical Fluid Chromatography-Tandem Mass Spectrometry.

Macrolides are a group of compounds used to treat bacterial infections in humans and animals. Their widespread use results in the contamination of the water environment, which, on the one hand, has a detrimental effect on aquatic organisms and, on the other hand, can lead to the emergence of resistant strains of microorganisms. All of the above determines the need for monitoring of these compounds in the environment, particularly, in water objects. Usually, the high-performance liquid chromatography combined with tandem mass spectrometry method is used to solve this problem, however, this work shows the possibility of using the supercritical fluid chromatography-tandem mass spectrometry method. An approach for the determination of four common macrolides, namely erythromycin, clarithromycin, midecamycin, and josamycin, was developed. The use of solid-phase extraction allowed to achieve limits of quantification at 0.57-6.8ng/L. The presented approach was validated and tested on a real object-a sample of municipal wastewater.

Characterization of a Soft Ionization by Chemical Reaction in Transfer Ion Source Hyphenated With Supercritical Fluid Chromatography-Tandem Mass Spectrometry.

A commercially available dielectric barrier discharge ionization (DBDI) source was tested with supercritical fluid chromatography-mass spectrometry (SFC-MS). The compound mixture investigated comprised caffeine, theobromine, theophylline, uracil, testosterone, and pyrene, diluted in methanol. Dynamic response ranges were evaluated with multiple injections at different concentrations. Precision studies demonstrated the robustness and sensitivity of the ionization source across a concentration range of 10-1000ng/mL. Results from this experiment showed linear regression of 0.99 or greater for all analytes tested over the range with a relative standard deviation (RSD) of less than 10% down to 10ng/mL for all analytes except theobromine, which had an RSD of less than 10% down to 25ng/mL. Notably, this study marks the first investigation of sensitivity for coupling a commercial DBDI source with SFC; a limit of detection less than 1ng/mL was achieved for all compounds. This study demonstrates chromatographic separation by SFC and MS analysis for compounds that ionize poorly using traditional atmospheric pressure ionization, such as polycyclic aromatic hydrocarbons. Combining SFC with the DBDI source opens promising avenues for analyzing compounds that were previously challenging to characterize with standard atmospheric pressure ionization techniques.

Quantitative determination of R/S-methadone in human serum using ultra-high performance supercritical fluid chromatography-tandem mass spectrometry: A method for routine use.

Methadone has two enantiomers, which exhibit differences in pharmacological effects, with R-methadone being the active and S-methadone the inactive enantiomer. A robust, simple and rapid method for chiral separation of the two enantiomers in serum samples using ultra-high performance supercritical fluid chromatography-tandem mass spectrometry (UHPSFC-MSMS) has been developed and validated. Enantiomeric separation was achieved using a Chiralpak IH-3 column with a mobile phase consisting of CO2 and 30mM ammonium acetate in methanol/water (98/2, v/v). Runtime was 4minutes. Sample preparation was semi-automated using a Hamilton ML Star robot with protein precipitation, and phospholipid removal was carried out using a Waters OSTRO™ 96-well plate. The calibration range was 50.0-1,500 nM for each enantiomer. The between-assay relative standard deviations were in the range of 1.2-3.6%. Matrix effects ranged from 99% to 115% corrected with internal standard. The method has been implemented in our laboratory and has proven to be a robust and reliable method for determining the ratio of R/S-methadone in authentic patient samples.

Evaluation of supercritical fluid chromatography coupled to tandem mass spectrometry for the analysis of pesticide residues in grain.

A supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS) technique was developed for the rapid and simultaneous detection of nine pesticides (carbendazim, isoprocarb, paclobutrazol, isoprothiolane, flusilazole, quinalphos, piperonylbutoxide, propargite, and bioresmethrin) in rice, wheat, and maize. The cereal samples were extracted with a solution of 0.5% acetic acid in acetonitrile and purified using quick, easy, cheap, effective, rugged, and safe method. The samples were characterized using multi-reaction monitoring and quantified with the external standard method. Excellent linearities (R2 >0.9991) and limits of quantification (0.4-40.0μg/kg) were established for all nine pesticides. Satisfactory pesticide recovery rates (62.2%-107.4%) were obtained at three standard concentrations (50, 100, and 200μg/kg), with relative standard deviations in the range of 2.1%-14.3%. The results confirmed that the proposed method was suitable for the routine detection of these pesticides in grain samples. Compared with high-performance liquid chromatography-MS/MS, the overall test run time and the amount of solvent required were reduced by 66% and 90%, respectively, when SFC-MS/MS was applied. Therefore, the use of SFC-MS/MS permits a shorter run time and affords greater analytical efficiency, such that it is both economical and environmentally sustainable.

Stereoselective bioaccumulation and dissipation of four stereoisomers of cyproconazole in earthworm-soil microcosm

Cyproconazole is a representative and widely used triazole fungicide with four stereoisomers, which will bring some risks to non-target organisms. A fast analytical method on supercritical fluid chromatography-tandem mass spectrometry was established in 4 min, and the environmental hazards of chiral cyproconazole were studied in earthworm-soil microcosm, including stereoselective bioaccumulation and dissipation. In the process of bioaccumulation, the concentrations of cyproconazole stereoisomers in earthworms showed a trend of increasing first and then reaching a stable state at 6 mg/kg treatment, which was different from those at 0.6 mg/kg treatment (decease-increase-equilibrium). The concentration order was (2S,3R)- > (2S,3S)- > (2R,3R)- > (2R,3S)-cyproconazole and (2S,3S)- ≈ (2S,3R)- > (2R,3R)- > (2R,3S)-cyproconazole at 6 and 0.6 mg/kg treatments, respectively. The bioaccumulation factor (BAF) values were in the range of 0.018–0.55, showing weakly relative accumulation capacity. The dissipation of cyproconazole stereoisomers in artificial soil accorded with the first-order kinetics equation, and the half-lives were 20.1–23.6 and 7.66–8.28 days at 6 and 0.6 mg/kg treatments, respectively, without stereoselectivity and diastereoselectivity. In earthworms, the dissipation half-lives were 5.81–6.01 days with the preferential dissipation of (2R,3R)-cyproconazole. The study would help with the rational uses and risk assessments of cyproconazole.

Novel approach to supercritical fluid chromatography-mass spectrometry analysis of metal ions using EDTA complexation

BackgroundReliable methods enabling detection of metal ions, and especially heavy metals, in different matrices are necessary in various fields such as ecology, pharmaceuticals and toxicology. As some of the currently used methods suffer from spectral and chemical interferences, this study investigates the applicability of SFC-MS/MS for the determination of metal ions. ResultsEffective novel approaches for metal ion analysis using CO2-based mobile phase were developed using three ligands forming metal complexes. As metal-EDTA complexes are prepared by simple addition of EDTA to the solution containing metal ions, this approach to metal ion analysis does not require laborious synthesis and isolation of solid metal-complexes. Besides, two other approaches using diethyldithiocarbamate and acetylacetonate as ligands were compared. Metal complexes of Cu, Co, Cr, Fe, Al, Mn, and Zn with all 3 ligands were synthesized and their identity was confirmed by high-resolution mass spectrometry (HRMS). The suitability of the three developed UHPSFC-MS/MS methods was examined using the determination of calibration range and repeatability of injections. Moreover, the universality of the developed UHPSFC-MS/MS method for the determination of metal-EDTA complexes was proved by analyzing Ni, Bi and Pb as additional metal ions. Significance and noveltyThis study demonstrates the extended range of applicability for SFC based separations. For the first time, the possibility to analyze metal complexes with EDTA using a fast and reliable ultra-high performance supercritical fluid chromatography-tandem mass spectrometry (UHPSFC-MS/MS) method is reported. The three developed UHPSFC-MS/MS methods are able to separate DDC, acac, and EDTA complexes of various metals very efficiently (total cycle times of 5, 2, and 3 min, respectively). They offer a fast and green alternative to chromatographic methods commonly used for metal ion analysis.

Determination of Oxygen Heterocyclic Compounds in Foods Using Supercritical Fluid Chromatography-Tandem Mass Spectrometry.

The aim of this research was to determine oxygen heterocyclic compounds in twenty-six Citrus- and cinnamon-flavoured foods using supercritical fluid chromatography in combination with triple-quadrupole mass spectrometry (SFC-QqQ-MS). According to the authors' knowledge, this is the first report on the determination of these molecules in foods by means of the SFC-QqQ-MS technique. The analytical technique normally used for their determination in foods is liquid chromatography coupled with a photodiode array detector. However, supercritical fluid chromatography is proving to be a valid alternative approach to investigating coumarins, furocoumarins and polymethoxyflavones. According to the results presented herein, each sample analysed showed the presence of molecules of interest. Coumarin was found in all the cinnamon-flavoured samples analysed in a low concentration. The presence of oxygen heterocyclic compounds in all the Citrus-flavoured samples, according to the label, comfirmed that the foods selected for this research article were prepared with Citrus fruits. Among the samples analysed, mandarin juice was the richest in bioactive compounds, representing a good source of polymethoxyflavones in a diet.

Comparison of supercritical fluid chromatography-tandem mass spectrometry and liquid chromatography-tandem mass spectrometry for the stereoselective analysis of chlorfenvinphos and dimethylvinphos in tobacco.

This study used reversed-phase liquid chromatography-tandem mass spectrometry and supercritical fluid chromatography-tandem mass spectrometry for determination of the stereoisomers of chlorfenvinphos and dimethylvinphos in tobacco. Tobacco samples were extracted and purified with a modified quick, easy, cheap, effective, rugged, and safe technique using spherical carbon. The performance of both methodologies was comprehensively compared in terms of methods validation parameters (separation efficiency, linearity, selectivity, recovery, repeatability, sensitivity, matrix effect, etc.). Under optimized conditions, the calibration curves of the stereoisomers of chlorfenvinphos and dimethylvinphos in the range of 10-500ng/mL showed excellent linearity with R2 ≥ 0.997 in both methods. The adequate recoveries of analytes from three different spiked tobaccos were obtained using reversed-phase liquid chromatography-tandem mass spectrometry (86.1-95.7%) as well as supercritical fluid chromatography-tandem mass spectrometry (86.5-94.0%). The relative standard deviations for spiked samples were all below 7.0%. Compared with supercritical fluid chromatography-tandem mass spectrometry, lower matrix effects and LODs can be obtained in reversed-phase liquid chromatography-tandem mass spectrometry.

Development and application of a supercritical fluid chromatography-tandem mass spectrometry method for the simultaneous determination of pantoprazole enantiomers in rat plasma

The study aimed to develop a supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS) method for the enantioselective separation and quantification of pantoprazole in rat plasma. The method utilized protein precipitation with acetonitrile to prepare plasma samples. SFC was performed on an Acquity ultra-performance convergence chromatography (UPCC) system. An Acquity UPCC Trefoil™ CEL2 column was employed for enantioseparation, and isocratic elution was achieved using a mobile phase consisting of CO2-methanol (81:19, v/v). The detection of pantoprazole enantiomers and the internal standard phenacetin was carried out using a Xevo TQD triple quadrupole mass spectrometer in selected reaction monitoring mode with positive electrospray ionization. The developed method successfully achieved full separation of S- and R-pantoprazole enantiomers. Calibration curves were linear in the concentration range of 10–5000 ng/mL for both S- and R-pantoprazole. The method met the acceptance criteria for carry-over, accuracy, precision, extraction recovery, matrix effect, stability, and dilution integrity, demonstrating its stereoselectivity, sensitivity, accuracy, precision, and reliability. The validated method was then applied to investigate the toxicokinetics of racemic pantoprazole and evaluate the potential of chiral inversion between S- and R-pantoprazole in rats. Following oral administration of 200 mg/kg racemic pantoprazole once daily for 90 consecutive days, there was no significant difference in toxicokinetic parameters between the two enantiomers on both day 1 (single-dose experiment) and day 90 (multiple-dose experiment). This suggests that there is no enantioselectivity in the toxicokinetic behaviors of racemic pantoprazole in rat plasma. Additionally, bidirectional chiral inversion between S- and R-pantoprazole in plasma was observed after single-dose and multiple-dose oral administrations of S-pantoprazole (20, 100, and 200 mg/kg) or R-pantoprazole (200 mg/kg) to rats. Compared to single and low doses, there was no accumulation of each enantiomer in rat plasma following repeated and escalated dosing. Overall, this study provided the first report on the toxicokinetics of pantoprazole in rat plasma and presented the first SFC-MS/MS method for the simultaneous quantification of pantoprazole enantiomers in rat plasma.

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.

Fabrication of magnetic magnesium oxide cleanup adsorbent for high-throughput pesticides residue analysis coupled with supercritical fluid chromatography-tandem mass spectrometry

A rapid and accurate analytical method was established for multiple pesticide residues in complex matrices based on magnetic dispersive solid phase extraction (d-SPE) and supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS). To develop an efficient magnetic d-SPE method, magnetic adsorbent modified with magnesium oxide (Fe3O4–MgO) was prepared via layer-by-layer modification and used as cleanup adsorbent for removal of interferences that contain a large number of hydroxyl or carboxyl groups in the complex matrix. The obtained Fe3O4–MgO coupled with 3-(N,N-Diethylamino)-propyltrimethoxysilane (PSA) and octadecyl (C18) were used as d-SPE purification adsorbents and their dosages were systematically optimized with Paeoniae radix alba as the matrix model. Combined with SFC-MS/MS, rapid and accurate determination of 126 pesticide residues in the complex matrix was achieved. Further systematic method validation showed good linearity, satisfactory recovery, and wide applicability. The average recoveries of the pesticides at 20, 50, 80, and 200 μg kg−1 were 110, 105, 108, and 109%, respectively. The proposed method was applied to complex medicinal and edible root plants, such as Puerariae lobate radix, Platycodonis radix, Polygonati odorati rhizoma, Glycyrrhizae radix, and Codonopsis radix. The average recoveries of the pesticides at 80 μg kg−1 in these matrices were 106, 106, 105, 103, and 105%, respectively with an average relative standard deviation range of 8.24–10.2%. The results demonstrated the feasibility and wide matrix applicability of the proposed method, which is promising for pesticide residue analysis in complex samples.

Analysis of 10 Phthalate Esters in Water-Based Adhesives by Supercritical Fluid Chromatography-Tandem Mass Spectrometry Combined With Spherical Carbons-Based Dispersion Solid-Phase Extraction.

Spherical carbons have a porous structure and large surface area for adsorption of macromolecules in water-based adhesives. Supercritical fluid chromatography (SFC) can improve selectivity and obtain better separation for phthalate esters (PAEs). The aim of this study was to develop a simple and green method for the simultaneous determination of 10 PAEs in water-based adhesives using SFC-tandem mass spectrometry with dispersion solid-phase extraction by spherical carbons. Separation of PAEs on a Viridis HSS C18 SB column and the parameters affecting the extraction were evaluated. Good accuracy and precision were obtained with the recoveries at 0.5, 2.0, and 10.0 mg/kg ranging from 82.9 to 99.5% and the intra- and inter-day precision less than 7%. The method had excellent sensitivity with limits of detection in the range of 0.015-0.029 mg/kg. In the 10-500 ng/mL concentration range, the linear correlation coefficients of all compounds were between 0.9975 and 0.9995. The method was applied to the determination of 10 PAEs in actual samples. This method is simple and rapid with low solvent consumption and high extraction efficiency. When applied to the determination of PAEs in actual samples, the method is sensitive and accurate and can meet the batch processing requirements for trace PAEs in water-based adhesives. PAEs in water-based adhesives can be determined using inexpensive materials and simple procedures with SFC.

Analysis of doping control samples using supercritical fluid chromatography-tandem mass spectrometry: Ready for routine use.

Supercritical fluid chromatography is proving to be a good separation and sample preparation tool for various analytical applications and, as such, has gained the attention of the anti-doping community. Here, the applicability of supercritical fluid chromatography hyphenated to tandem mass spectrometry for routine doping control analysis was tested. A multi-analyte method was developed to cover 197 drugs and metabolites that are prohibited in sport. More than 1000 samples were analyzed by applying a "dilute and inject" approach after hydrolysis of glucuronide metabolites. Additionally, a comparison with routinely used liquid chromatography-mass spectrometry was performed with 250 of the 1000 samples and a number of past positive anti-doping samples. It revealed some features where supercritical fluid chromatography-tandem mass spectrometry was found to be complementary or advantageous to liquid chromatography-mass spectrometry for anti-doping purposes, such as better retention of analytes that are poorly retained in reversed-phase liquid chromatography. Our results suggest that supercritical fluid chromatography-tandem mass spectrometry is sensitive (limit of detection <50% relevant minimum required performance level required by the World Anti-Doping Agency for anti-doping analysis), reproducible, robust, precise (analytes of interest area coefficient of variation <5%; retention time difference coefficient of variation <1%) and complementary to existing techniques currently used for routine analysis in the World Anti-Doping Agency accredited laboratories.

Stereoselective analysis of chiral succinate dehydrogenase inhibitors (SDHIs) in foods of plant origin and animal origin by supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS)

The stereoisomers of chiral SDHIs were prepared using Autoprep HPLC and chiral columns. The method of combining theoretical calculation with experimental determination was used to confirm the absolute configuration of stereoisomer. SFC-MS/MS and four kinds of chiral columns were used to separate the eight chiral SDHIs, and they could be separated simultaneously using OD-3 column in 6.5 min. The integrated QuEChERS strategy was used to analyse the chiral SDHIs in foods of plant and animal origin, and the average recoveries ranged from 71 % to 119 % with RSD ≤ 18 %, and the LOQ was 1 ng/g. There were 99.2 % and 63.6 % matrix effects were in the range of 0.8–1.2 in foods of plant and animal origin, respectively, showing weak matrix effects. The study provided methods for monitoring chiral SDHIs stereoisomers residues, which were crucial for stereoselective evaluations and improving risk assessments.

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.

Study on the enantioselective bioaccumulation and dissipation of uniconazole enantiomers in earthworm-soil microcosm through supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS).

In this work, the enantioselective bioaccumulation and dissipation of uniconazole enantiomers in earthworm-soil microcosm were studied. A fast enantioseparation method of uniconazole through supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS) was established. The CHIRALCEL OZ-3 column and a mixture of CO2 and methanol (80:20, v/v) were used within 1.0min to separate uniconazole enantiomers. The recoveries of uniconazole enantiomers in earthworm and soil samples ranged from 83.3 to 113%, and the intra-day and inter-day relative standard deviation values were lower than 11%. In earthworms, the bioaccumulation concentrations of uniconazole enantiomers increased with time and reached the maximum on the 7th day and then decreased. The elimination of uniconazole enantiomers in earthworms followed the first-order kinetics equation, and the elimination half-lives were approximately 7days. In artificial soil, the dissipation of uniconazole enantiomers was slow, and the dissipation half-lives were both 25.7days. No enantioselectivity occurred in the earthworm-soil microcosm. These results may reduce the uncertainty of environmental risk assessment for uniconazole.

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.

Ultra-high performance supercritical fluid chromatography-tandem mass spectrometry method for simultaneous determination of atorvastatin, 2-hydroxy atorvastatin, and tangeretin in rat plasma and its application to the pharmacokinetic study.

Recent studies strongly suggest that atorvastatin combination therapy with tangeretin could be beneficial in the treatment of hyperlipidemia. This study aimed to investigate the pharmacokinetic interactions among atorvastatin, its active metabolite 2-hydroxy atorvastatin, and tangeretin after oral administration of atorvastatin with tangeretin in rats. A rapid, selective, and sensitive assay was developed and validated based on ultra-high performance supercritical fluid chromatography-tandem mass spectrometry for the simultaneous measurement of atorvastatin, 2-hydroxy atorvastatin, and tangeretin concentrations in rat plasma. Chromatographic separation of the analytes was conducted on an ACQUITY Torus 1-AA column in gradient elution mode. The mass transition ion pairs were m/z 559.0→440.0 for atorvastatin, m/z 575.2→440.0 for 2-hydroxy atorvastatin, m/z 373.0→358.1 for tangeretin, and m/z 254.8→136.7 for daidzein (internal standard). Calibration curves showed good linear correlations at the following concentration range: 1-400 (r = 0.9952), 1-400 (r = 0.9980), and 3-1200 (r = 0.9945) for atorvastatin, 2-hydroxy atorvastatin, and tangeretin, respectively. The method was fully validated and satisfied the acceptance criteria recommended by the United States Food and Drug Administration. Finally, it was successfully applied in a pharmacokinetic study in rats to evaluate the pharmacokinetic behavior of atorvastatin, 2-hydroxy atorvastatin, and tangeretin.

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.