Chromatography-atmospheric Pressure Chemical Ionization Mass Research Articles

Rapid Quantification of Endogenous Steroids in Human Serum Using Leidenfrost Effect-Assisted Thermal Desorption Atmospheric Pressure Photoionization Orbitrap Mass Spectrometry.

Unconjugated sex steroids in human serum play a crucial role in physiological and pathological studies and are frequently considered as biomarkers in clinical diagnosis. Because of their low polarity, poor volatility, and low concentration, the rapid and highly sensitive analysis of sex steroids in real serum matrix by ambient mass spectrometry is still challenging. Here, Leidenfrost effect-assisted thermal desorption atmospheric pressure photoionization orbitrap mass spectrometry (LETD-APPI-MS) was developed and applied to quantify free sex steroids in human serum without derivatization and chromatography separation within a few minutes. The concentration of target analyte could be increased by approximately two orders during the LETD process. The limit of quantifications and detections of endogenous sex steroids in human serum were measured at the ppt level. In contrast with commonly used immunoassays in clinical laboratories, LETD-APPI-MS enables the accurate measurements of multiple free sex steroids without the interference of cross-reactions. The endogenous sex steroids of 38 female serums at four physiological stages during pregnancy were rapidly tested by LETD-APPI-MS, whose results were highly consistent with that using liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC-APCI-MS), indicating LETD-APPI-MS has a strong clinical application potential in steroid analysis.

Contrastive analysis of lipid composition and thermal and crystallization behavior of olein/stearin fractionated by novel layer melt crystallization from palm oil.

Melt crystallization is typically recognized as a highly efficient and green method for oil fractionation. This work concentrated on novel layer melt crystallization for preparing desirable olein and stearin products from palm oil and the evaluation of fraction quality. Layer melt crystallization was performed at various temperatures and the effects on fractions were evaluated using iodine value (IV), solid fat content (SFC) and melting point. The lipid composition, thermal and crystallization properties, and phase behaviors of the final optimized fractions were determined using gas chromatography, high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry and differential scanning calorimetry. Increasing crystallization tube temperatures under the same jacket temperature increased the melting point and SFC, while decreasing the IV of the olein product. Opposite results were observed for the stearin product. Major fatty acids in fractions were determined as palmitic acid and oleic acid. 1,2-Dioleoyl-3-palmitoylglycerol and 1,3-dipalmitoyl-2-oleoylglycerol were identified as the main triacylglycerols in olein and stearin fractions, respectively. A critical effect of crystallization temperature on co-crystallization of oleins and stearins was revealed. A transition from plate-like crystal growth to spherulitic growth with spontaneous nucleation was indicated in palm oil and stearin fractions with increasing crystallization temperature. As for olein fractions, a temperature increase resulted in heterogeneous nucleation from instantaneous nucleation. Novel layer melt crystallization was successfully applied and optimized for fractionating palm oil. The composition and property changes of obtained fractions were analyzed and explained at both macroscopic and microscopic levels. © 2021 Society of Chemical Industry.

Sebomic identification of sex- and ethnicity-specific variations in residual skin surface components (RSSC) for bio-monitoring or forensic applications

Background“Residual skin surface components” (RSSC) is the collective term used for the superficial layer of sebum, residue of sweat, small quantities of intercellular lipids and components of natural moisturising factor present on the skin surface. Potential applications of RSSC include use as a sampling matrix for identifying biomarkers of disease, environmental exposure monitoring, and forensics (retrospective identification of exposure to toxic chemicals). However, it is essential to first define the composition of “normal” RSSC. Therefore, the aim of the current study was to characterise RSSC to determine commonalities and differences in RSSC composition in relation to sex and ethnicity.MethodsSamples of RSSC were acquired from volunteers using a previously validated method and analysed by high-pressure liquid chromatography–atmospheric pressure chemical ionisation–mass spectrometry (HPLC-APCI-MS). The resulting data underwent sebomic analysis.ResultsThe composition and abundance of RSSC components varied according to sex and ethnicity. The normalised abundance of free fatty acids, wax esters, diglycerides and triglycerides was significantly higher in males than females. Ethnicity-specific differences were observed in free fatty acids and a diglyceride.ConclusionsThe HPLC-APCI-MS method developed in this study was successfully used to analyse the normal composition of RSSC. Compositional differences in the RSSC can be attributed to sex and ethnicity and may reflect underlying factors such as diet, hormonal levels and enzyme expression.

UHPLC-APCI-MS Profiling of Triacylglycerols in Vegetable Oils—Application to the Analysis of Four North African Sesame Seed Varieties

Here, we describe an isocratic ultra-high performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (UHPLC-APCI-MS) method for the characterization and the quantification of triacylglycerols (TAGs) in vegetable oils. The linearity and the precision of the method are validated, and the limits of detection are estimated. The APCI-MS detector shows a linear response from 1 to 1000 μg/mL, and the correlation coefficients (r2) obtained for standards TAGs exceed 0.995. As compared to previous methods, the proposed method is rapid, simpler, and more sensitive. The limits of detection estimated for several standard TAGs varies from 0.03 to 0.05 injected nanograms (6 to 10 μg/L). The application of the proposed method to the analysis of sesame oil allowed us to unambiguously identify 30 TAG species whereas previous methods have detected 19 TAGs, at the most. Among vegetable oils, sesame oil became an attractive target for adulteration due to its high commercial value. Most of the previous studies of TAG profiles of sesame oils had been devoted to the analysis of sesame oils originating from Asia and some Sub-Saharan countries. To the best of our knowledge, the TAG profiles of sesame oils from other origins are still unknown. The results we obtained for the four Tunisian sesame seed varieties clearly indicate that sesame oils can be classified according to their equivalent carbon number differences (ΔECN) with some rules similar to those required for olive oils.

Separation and Identification of Odd Chain Triacylglycerols of the Protozoan Khawkinea quartana and the Mold Mortierella alpina Using LC-MS.

Liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC-MS/APCI) with reversed- and chiral phases was used for separation of triacylglycerols (TAG) from protozoan and mold. This study describes the separation and identification of odd numbered chains of regioisomers and enantiomers of triacylglycerols from different natural sources, i.e., the protozoan Khawkinea quartana and the mold Mortierella alpina. Using the above-mentioned separation methods and the synthesis of appropriate standards of TAG, we identified regioisomers and enantiomers of both even and odd numbered TAG. The biosynthesis of odd numbered TAG was found to be strictly stereospecific and to depend on the production microorganism, one enantiomer predominating in the protozoan and the other in the mold. It was proved that even numbered TAG are synthesized in a higher optical purity, which can be explained by a higher affinity of acyltransferases to the respective substrate, i.e., to even chain PUFA.

Quantitation of triacylglycerols in edible oils by off-line comprehensive two-dimensional liquid chromatography–atmospheric pressure chemical ionization mass spectrometry using a single column

In this investigation, off-line comprehensive two-dimensional liquid chromatography–atmospheric pressure chemical ionization mass spectrometry using a single column has been applied for the identification and quantification of triacylglycerols in edible oils. A novel mixed-mode phenyl-hexyl chromatographic column was employed in this off-line two-dimensional separation system. The phenyl-hexyl column combined the features of traditional C18 and silver-ion columns, which could provide hydrophobic interactions with triacylglycerols under acetonitrile conditions and can offer π–π interactions with triacylglycerols under methanol conditions. When compared with traditional off-line comprehensive two-dimensional liquid chromatography employing two different chromatographic columns (C18 and silver-ion column) and using elution solvents comprised of two phases (reversed-phase/normal-phase) for triacylglycerols separation, the novel off-line comprehensive two-dimensional liquid chromatography using a single column can be achieved by simply altering the mobile phase between acetonitrile and methanol, which exhibited a much higher selectivity for the separation of triacylglycerols with great efficiency and rapid speed. In addition, an approach based on the use of response factor with atmospheric pressure chemical ionization mass spectrometry has been developed for triacylglycerols quantification. Due to the differences between saturated and unsaturated acyl chains, the use of response factors significantly improves the quantitation of triacylglycerols. This two-dimensional liquid chromatography–mass spectrometry system was successfully applied for the profiling of triacylglycerols in soybean oils, peanut oils and lord oils. A total of 68 triacylglycerols including 40 triacylglycerols in soybean oils, 50 triacylglycerols in peanut oils and 44 triacylglycerols in lord oils have been identified and quantified. The liquid chromatography–mass spectrometry data were analyzed using principal component analysis. The results of the principal component analysis enabled a clear identification of different plant oils. By using this two-dimensional liquid chromatography–mass spectrometry system coupled with principal component analysis, adulterated soybean oils with 5% added lord oil and peanut oils with 5% added soybean oil can be clearly identified.

Separation of enantiomeric triacylglycerols by chiral-phase HPLC.

Liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC-MS/APCI) with a chiral phase was used for separation of triacylglycerols (TAG) obtained either by organic synthesis or isolated from different algal species. We present chromatographic characteristics and tandem mass spectra of enantiomers and positional isomers (regioisomers) of C16, C18 and C20 polyunsaturated fatty acids. The retention time was found to depend on the structure of the given TAG, increasing with increasing number of double bonds and decreasing with increasing number of the carbons in TAG, with the exception of dieicosapentaenoyl-palmitoyl-glycerols.

Identification of the protopanaxatriol synthase gene CYP6H for ginsenoside biosynthesis in Panax quinquefolius.

Panax quinquefolius is one of perennial herbs and well known for its outstanding pharmacological activity. Ginsenosides are thought to be the main active ingredients in P. quinquefolius and exist in many kinds of plant genus Panax (ginseng). Protopanaxatriol synthase, which is considered cytochrome P450 (CYP450) in ginsenoside biosynthesis pathway can convert protopanaxadiol into protopanaxatriol. However, the protopanaxatriol synthase gene in P. quinquefolius has not been identified. Here, we cloned and identified a protopanaxatriol synthase gene from P. quinquefolius (CYP6H, GenBank accession no. KC190491) at the first time, reverse transcription-PCR (RT-PCR) analysis showed no obvious transcription change of CYP6H in methyl jasmonate (MeJA)-induced hairy roots. Ectopic expression of CYP6H in Saccharomyces cerevisiae resulted in the production of protopanaxatriol with added exogenous protopanaxadiol and confirmed by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC/APCIMS). Moreover, high-performance liquid chromatography (HPLC) analysis shows that RNA interferences of CYP6H in transgenic hairy roots could increase the accumulation of protopanaxadiol-type ginsenosides and decrease the accumulation of protopanaxatriol-type ginsenosides, whereas the effect of overexpression CYP6H in transgenic hairy roots was contrary. Our study indicated that CYP6H is a gene encoding protopanaxadiol 6-hydroxylase which could convert protopanaxadiol into protopanaxatriol in P. quinquefolius ginsenoside biosynthesis, we also have confirmed the function of CYP6H on effect accumulation of ginsenosides.

The isolation and characterization of dammarenediol synthase gene from Panax quinquefolius and its heterologous co-expression with cytochrome P450 gene PqD12H in yeast.

Panax quinquefolius is one of perennial herbs and well known for its outstanding pharmacological activity. Ginsenosides are thought to be the main active ingredients in Panax quinquefolius and exist in many kinds of plant genus Panax (ginseng). Dammarenediol synthase, which is considered as a key enzyme in ginsenoside biosynthesis pathway can convert 2, 3-oxidosqualene into dammarenediol-II. However, the dammarenediol synthase gene in Panax quinquefolius has not been identified. Here, we cloned and identified a dammarenediol synthase gene from Panax quinquefolius (PqDS, GenBank accession No. KC316048) at the first time, and reverse transcription-PCR (RT-PCR) analysis also showed an obvious transcription increase of PqDS in the methyl jasmonate (MeJA)-induced hairy roots. Ectopic expression of PqDS in yeast resulted in the production of dammarenediol-II was confirmed by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC/APCIMS). Moreover, overexpression of PqDS in transgenic hairy roots could increase the transcription of gene PqDS and another P450 gene PqD12H (encoding protopanaxadiol synthase in Panax quinquefolius), the accumulation of ginsenosides also increased at the same time. In addition, both PqDS and PqD12H gene co-expressed in recombinant yeast result in the production of protopanaxadiol was detected by LC/APCIMS; this result also provides a new strategy for the abundant production of protopanaxadiol in vitro.

Comparative analysis of triacylglycerols from different Stichococcus strains by RP-HPLC/APCI-MS and chiral HPLC

The main analytical benefit of this study is the development of methods enabling a rapid determination of triacylglycerols (TAGs) in different strains of the green alga Stichococcus bacillaris and detailed identification and quan- tification of a complex mixture of natural TAGs by reversed phase liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (RP-HPLC/APCI-MS) and chi- ral HPLC. Neutral ion scan can readily identify, both qualita- tively and semiquantitatively, triacylglycerols containing 16:2 and16:3fattyacids(FAs)inthemolecule.Sincemoredetailed studies in this respect have been stymied by the shortage of 16:2 and 16:3 FAs, we decided to study the alga Stichococcus as a potential new biotechnological source of C16 polyunsat- urated fatty acids (PUFAs). We conclude that the genus Stichococcus is a major producer of C16 PUFAs mostly contained in TAGs. Further, the ratio of regioisomers and enantiomers of TAGs depends on the content of fatty acids; with lowering temperature, the proportion of the achiral TAG increases, and so does the enantiomer ratio from 1:1. Dienoic FAs preferentially esterify position sn-3, whereas in trienoic acids, it is position sn-1.

Accurate quantitation of pentaerythritol tetranitrate and its degradation products using liquid chromatography–atmospheric pressure chemical ionization–mass spectrometry

After an explosion of pentaerythritol tetranitrate (PETN), its degradation products pentaerythritol trinitrate (PETriN), dinitrate (PEDiN) and mononitrate (PEMN) were detected using liquid chromatography–atmospheric-pressure chemical-ionization–mass spectrometry (LC–APCI–MS). Discrimination between post-explosion and naturally degraded PETN could be achieved based on the relative amounts of the degradation products. This information can be used as evidence when investigating a possible relationship between a suspect and a post-explosion crime scene. The present work focuses on accurate quantitation of PETN and its degradation products, using PETriN, PEDiN and PEMN standards specifically synthesized for this purpose. With the use of these standards, the ionization behavior of these compounds was studied, and a quantitative method was developed. Quantitation of PETN and trace levels of its degradation products was shown to be possible with accuracy between 85.7% and 103.7% and a precision ranging from 1.3% to 11.5%. The custom-made standards resulted in a more robust and reliable method to discriminate between post-explosion and naturally-degraded PETN.

CHEMOMETRIC APPROACHES FOR THE CHARACTERIZATION OF TURKISH RHODODENDRON AND HONEYDEW HONEYS DEPENDING ON AMINO ACID COMPOSITION

The amino acid composition of twenty-five honey samples obtained from different regions of Turkey (rhododendron and honeydew honey) was determined by a liquid chromatography–atmospheric pressure chemical ionization–mass spectrometry (LC–APCI–MS). Principal component analysis (PCA) and hierarchical cluster analysis (HCA) were used to characterize and classify the honey samples. In rhododendron honeys, major amino acids were detected as aspartic acid, lysine, and arginine, for honeydew honeys, lysine, arginine, and histidine were found, following the proline. Five principal components (PCs) and four main clusters were obtained from the chemometric analysis that classifies the samples. Five PCs were detected to be explanatory of more than 79% of the total variance in the data matrix. It can be concluded that free amino acids are good indicators for determination of botanical origins and chemometric studies are highly useful for honey characterization.

Simultaneous pharmacokinetic assessment of cefadroxil and clavulanic acid in human plasma by LC–MS and its application to bioequivalence studies

A simple, rapid and selective liquid chromatography–atmospheric pressure chemical ionization–mass spectrometry (LC–APCI–MS) assay method has been developed and fully validated for the simultaneous quantification of cefadroxil (CF) and clavulanic acid (CA) in human plasma. Analytes and internal standard (IS) were extracted from human plasma by solid-phase extraction (SPE) technique using Sam prep (3mL, 100mg) extraction cartridge. The extracted samples were chromatographed on a reverse phase C18 column using a mixture of methanol: acetonitrile: 2mM ammonium acetate (pH 3.5) (25:25:50, v/v/v) as the mobile phase at a flow rate of 0.8mL/min. Quantification of the analytes and IS were carried out using single quadrupole LC–APCI–MS through selected-ion monitoring (SIM) at m/z 362 and m/z 198, for CF and CA, respectively. Method validation was performed as per the FDA guidelines and the results met the acceptance criteria. Plasma concentration of CF and CA followed by the oral administration of CF/CA (500/125mg) pill to healthy male volunteers (n=12) was measured. Area under plasma concentration–time curve from 0 to 12h (AUC0–12h) and 0h extrapolated to infinity (AUC0−∞) were calculated. The ratio of AUC0–12h/AUC0−∞ was found to be >85% for all the subjects, as recommended by the FDA guidelines.

Simultaneous quantification of vinblastine and desacetylvinblastine concentrations in canine plasma and urine samples using LC–APCI–MS/MS

A highly sensitive and specific liquid chromatography–atmospheric pressure chemical ionization–mass spectrometry (LC/APCI–MS/MS) method has been developed and validated for simultaneous quantification of vinblastine and its metabolite, desacetylvinblastine, in canine plasma and urine samples. Plasma and urine samples were processed by a solid phase extraction procedure. The optimal chromatographic behavior of these analytes was achieved on pentafluorophenyl (PFP) propyl analytical column (5μm, 50×2.1mm) under isocratic elution of 0.75mL/min with a mobile phase of 5mM ammonium acetate and methanol. The samples were analyzed in positive ion, multiple reaction monitoring mode. The calibration curves were linear over 0.125–2ng/mL (lower calibration curve); 2–100ng/mL (higher calibration curve) and 0.125–5ng/mL for vinblastine and desacetylvinblastine in plasma, and over 1–2000ng/mL and 0.5–100ng/mL for vinblastine and desacetylvinblastine in urine samples, respectively. The limits of quantitation of vinblastine and desacetylvinblastine were 0.125ng/mL in both matrices. The intra and interday accuracy was above 89% and precision below 8.6% for both analytes in both matrices. The developed method was successfully applied to ongoing in vivo vinblastine pharmacokinetic studies in dogs.

Triacylglyceride profiling in serum by silver ion high performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry

A simple and effective method was developed to investigate triacylglyceride (TAG) compounds in biological matrices using silver ion high performance liquid chromatography-mass spectrometry (Ag+ -HPLC-MS). The TAG compounds from mouse serum were extracted by a modified Folch method using the classical CHC13-MeOH solvent system. The extract was separated on a Varian ChromSpher 5 Lipids column with isocratic elution using acetonitrile-hexane (1: 99, v/v) as mobile phase at a flow rate of 0.75 mL/min, and detected by a mass spectrometer equipped with an atmospheric pressure chemical ionization (APCI) source in positive ion mode to acquire sufficient MS information of TAG compounds by enhanced mass spectrometry (EMS), enhanced product ion (EPI) and neutral loss (NL) scans. The identification of TAG compounds was based on their chromatographic behaviors and MS data. Besides, the NL scans of thirty fatty acids were performed to further validate the results. Finally, 66 TAG compounds as well as 5 cholesteryl ether (CE) compounds were obtained from the mouse serum extract. This method is simple, reproducible and also suitable for the analysis of TAG compounds in other samples.

Cytochrome P450 CYP716A53v2 Catalyzes the Formation of Protopanaxatriol from Protopanaxadiol During Ginsenoside Biosynthesis in Panax Ginseng

Ginseng (Panax ginseng C.A. Meyer) is one of the most popular medicinal herbs, and the root of this plant contains pharmacologically active components, called ginsenosides. Ginsenosides, a class of tetracyclic triterpene saponins, are synthesized from dammarenediol-II after hydroxylation by cytochrome P450 (CYP) and then glycosylation by a glycosyltransferase. Protopanaxadiol synthase, which is a CYP enzyme (CYP716A47) that catalyzes the hydroxylation of dammarenediol-II at the C-12 position to yield protopanaxadiol, was recently characterized. Here, we isolated two additional CYP716A subfamily genes (CYP716A52v2 and CYP716A53v2) and determined that the gene product of CYP716A53v2 is a protopanaxadiol 6-hydroxylase that catalyzes the formation of protopanaxatriol from protopanaxadiol during ginsenoside biosynthesis in P. ginseng. Both CYP716A47 and CYP716A53v2 mRNAs accumulated ubiquitously in all organs of ginseng plants. In contrast, CYP716A52v2 mRNA accumulated only in the rhizome. Methyl jasmonate (MeJA) treatment resulted in the obvious accumulation of CYP716A47 mRNA in adventitious roots. However, neither CYP716A52v2 nor CYP716A53v2 mRNA was affected by MeJA treatment during the entire culture period. The ectopic expression of CYP716A53v2 in recombinant WAT21 yeast resulted in protopanaxatriol production after protopanaxadiol was added to the culture medium. In vitro enzymatic activity assays revealed that CYP716A53v2 catalyzed the oxidation of protopanaxadiol to produce protopanaxatriol. The chemical structures of the protopanaxatriol products were confirmed using liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC/APCIMS). Our results indicate that the gene product of CYP716A53v2 is a protopanaxadiol 6-hydroxylase that produces protopanaxatriol from protopanaxadiol, which is an important step in the formation of dammarane-type triterpene aglycones in ginseng saponin biosynthesis.

Regioisomer separation and identification of triacylglycerols containing vaccenic and oleic acids, and α- and γ-linolenic acids, in thermophilic cyanobacteria Mastigocladus laminosus and Tolypothrix sp.

Reversed phase liquid chromatography–atmospheric pressure chemical ionization mass spectrometry (RP-HPLC/APCI-MS) was used for direct analysis of triacylglycerols (TAGs) from different strains of the cyanobacteria Mastigocladus laminosus, Tolypothrix cf. tenuis and Tolypothrix distorta. This technique enabled us to identify and quantify the specific molecular species of TAGs directly from lipid extracts of the cyanobacteria. The regioisomeric series of TAGs having α-linolenic and γ-linolenic and also oleic and cis-vaccenic acids were separated by RP-HPLC and identified by APCI-MS. M. laminosus produced only a few molecular species of TAGs, including both isomers of octadecenoic (oleic and vaccenic) acid, while T. distorta contained tens of molecular species of TAGs having FAs with up to four double bonds (stearidonic acid and including also its positional isomer, i.e. 3,6,9,12-octadecatetraenoic acid) and both positional isomers (α and γ) of linolenic acids. Individual strains of both cyanobacteria exhibited different contents of polyunsaturated fatty acids (Tolypothrix sp.) and different distribution of positional isomers of monoenoic fatty acids in TAGs (M. laminosus).

The Cyt P450 Enzyme CYP716A47 Catalyzes the Formation of Protopanaxadiol from Dammarenediol-II During Ginsenoside Biosynthesis in Panax ginseng

Ginseng (Panax ginseng C.A. Meyer) is one of the most popular medicinal herbs and contains pharmacologically active components, ginsenosides, in its roots. Ginsenosides, a class of tetracyclic triterpene saponins, are thought to be synthesized from dammarenediol-II after hydroxylation by the Cyt P450 (CYP) enzyme and then glycosylation by glycosyltransferase (GT). However, no genes encoding the hydroxylation and glycosylation in ginsenoside biosynthesis have been identified. Here, we identify protopanaxadiol synthase, which is a CYP enzyme (CYP716A47), to be involved in the hydroxylation of dammarenediol-II at the C-12 position to yield protopanaxadiol. Nine putative full CYP sequences were isolated from the expressed sequence tags (ESTs) of methyl jasmonate (MeJA)-treated adventitious ginseng roots. The CYP716A47 gene product was selected as the putative protopanaxadiol synthase because this gene was transcriptionally activated not only by MeJA treatment but also in transgenic ginseng that overexpresses squalene synthase and overproduces ginsenosides. In vitro enzymatic activity assays revealed that CYP716A47 catalyzed the oxidation of dammarenediol-II to produce protopanaxadiol. Ectopic expression of CYP716A47 in recombinant WAT21 yeasts that were fed dammarenediol-II yielded protopanaxadiol. Furthermore, co-expression of the dammarenediol synthase gene (PgDDS) and CYP716A47 in yeast yielded protopanaxadiol without adding dammarenediol-II. The chemical structures of the protopanaxadiol products from dammarenediol-II were confirmed using liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC/APCIMS). Thus, CYP716A47 is a dammarenediol 12-hydroxylase that produces protopanaxadiol from dammarenediol-II.

The genus Dracunculus – A source of triacylglycerols containing odd-numbered ω-phenyl fatty acids

Reversed phase liquid chromatography–atmospheric pressure chemical ionization mass spectrometry (RP-HPLC/MS-APCI) was used to identify and quantify triacylglycerols (TAGs) having odd-numbered ω-phenylalkanoic acids from seeds of the flower plant Dracunculus vulgaris, and TAGs from the bacterium Rhodococcus erythropolis prepared by precursor directed biosynthesis from phenylalanine and having the corresponding even-numbered ω-phenylalkanoic acids. Model compounds, which are not commercially available, were prepared by organic synthesis and this allowed us to extend the number of identified natural TAGs to nearly 140 molecular species. Both synthetic and natural compounds containing ω-phenylalkanoic acids were found to have antioxidant and free radical scavenging properties.

Highly sensitive, quick and simple quantification method for mono and disaccharides in aqueous media using liquid chromatography–atmospheric pressure chemical ionization–mass spectrometry (LC–APCI–MS)

A highly sensitive, rapid LC–APCI–MS method for identification and quantification of mono and disaccharides in simple or complex aqueous phase has been developed. This original method is easy to use, no derivation and no post-column injection are needed. The separation is performed with a hydrophilic amino interaction (HILIC) column allowing high-throughput analysis with analysis times of 15 min for monosaccharides to 22 min for disaccharides. The development of the method carried out with 9 standard saccharides allowed to point out a dynamic range from 0.1–25.6 to 1–256 μg mL −1 depending on the considered sugar. Next, the method was validated on saccharides at known concentrations in water and on 2 real samples: orange juice and aqueous phase obtained after enzymatic hydrolysis of sunflower seeds.