Search

Naphthenic acids comprise one of the most toxic compounds of the produced water released from offshore oil platforms. Therefore, developing and applying faster, simpler, and more efficient analytical methods for analyzing naphthenic acids are urgently needed. Electromembrane extraction (EME) uses the electrokinetic migration of target ions through a porous membrane. Herein, the EME method was applied to extract naphthenic acids from produced water. The EME method was optimized, and the optimal conditions encompassed decanol as the organic solvent, the sample with pH 10.0, 5 min of extraction at 200 V, and the ratio 4:1 (borate buffer/matrix, v/v). Electrochemical impedance spectroscopy confirmed charged species' migration from produced water through the EME. Subsequently, all extracts were analyzed by ultra-high-resolution mass spectrometry. The EME efficiency was assessed by comparing the extraction results to the liquid-liquid extraction (LLE) method results. Analytical results showed good linearity for both solvent and matrix curves (

Stingless bee honey (SBH) is gaining attention due to its nutritional, sensorial, and medicinal characteristics. This study focuses on the combination of physicochemical properties, antioxidant capacity, mineral profile, and mass spectrometry-based fingerprints, using a chemometric approach to differentiate SBH (n = 18) from three different Brazilian biogeographical zones (Caatinga, Cerrado, and Atlantic Forest). The physicochemical properties of SBH varied, resulting in a wide range of water activity, moisture, total soluble solids, pH, and total and free acidity. The Caatinga honey showed the highest and the lowest contents of phenolics and flavonoids, respectively. The antioxidant free-radical scavenging assays demonstrated that the Brazilian SBH has a high antioxidant potential. The mineral profile of honey samples from the Atlantic Forest revealed higher contents of Ca and Fe while the Cerrado and Caatinga honey showed the highest P contents. Partial Least-Squares Discriminant Analysis (PLS-DA) analysis separated the samples into three groups based on the biogeographical zones of harvest. The main separation factors between groups were the m/z 326 ion and the Fe content. Univariate analysis confirmed that Fe content is important for SBH discrimination. The present results indicate that the origin of SBH can be determined on the basis of mineral profile, especially Fe content.

This work describes the surface coating of wooden toothpicks with amino groups (NH

Graphical abstract The novel coronavirus SARS-CoV-2 has been affecting the world, causing severe pneumonia and acute respiratory syndrome, leading people to death. Therefore, the search for anti-SARS-CoV-2 compounds is pivotal for public health. Natural products may present sources of bioactive compounds; among them, flavonoids are known in literature for their antiviral activity. Siparuna species are used in Brazilian folk medicine for the treatment of colds and flu. This work describes the isolation of 3,3′,4′-tri-O-methyl-quercetin, 3,7,3′,4′-tetra-O-methyl-quercetin (retusin), and 3,7-di-O-methyl-kaempferol (kumatakenin) from the dichloromethane extract of leaves of Siparuna cristata (Poepp. & Endl.) A.DC., Siparunaceae, using high-speed countercurrent chromatography in addition to the investigation of their inhibitory effect against SARS-CoV-2 viral replication. Retusin and kumatakenin inhibited SARS-CoV-2 replication in Vero E6 and Calu-3 cells, with a selective index greater than lopinavir/ritonavir and chloroquine, used as control. Flavonoids and their derivatives may stand for target compounds to be tested in future clinical trials to enrich the drug arsenal against coronavirus infections. Supplementary Information The online version contains supplementary material available at 10.1007/s43450-021-00162-5.

The proteomic data presented in this article are associated with the research article entitled “Longissimus dorsi muscle label-free quantitative proteomic reveals biological mechanisms associated with intramuscular fat deposition” published in Journal of Proteomics [1] . In this article, we characterized the proteomic profile of bovine Longissimus dorsi muscle from Nelore steers and identified differentially abundant proteins associated with the intramuscular fat (IMF) content. An integrated transcriptome-assisted label-free quantitative proteomic approach by High Definition Mass Spectrometry (HDMSE) was employed to identify and quantify the proteins. A functional enrichment analysis using the differentially abundant proteins list was performed to understand the biological processes involved in IMF deposition. Moreover, to explore and clarify the biological mechanisms that influence IMF content, the mRNA data for the same trait from Cesar and collaborators [2] obtained by RNA-sequencing technology was compared with proteomic data. The mRNA data is deposited in the European Nucleotide Archive (ENA) repository (EMBL-EBI), under accession PRJEB13188.

The influence of in vitro maturation (IVM) in oocytes is still not totally understood. The aim of this study was to determine the influence of IVM on the metabolism and homeostasis of bovine cumulus-oocyte complexes. In the present study, we demonstrated that IVM leads to accumulation of neutral lipids associated with differential levels of the mono-, di- and triacylglycerols in both cumulus cells and oocytes. We observed that in vitro-matured oocytes exhibited decreased glutathione and reactive oxygen species levels and a lower ATP/ADP ratio when compared to in vivo-matured oocytes, with no significant differences in metabolism and stress-related mRNA or miRNA levels. Moreover, in addition to an increase in lipids in in vitro-matured cumulus cells, fatty acid synthesis and accumulation as well as glycolysis pathway genes were upregulated, whereas those affiliated with the β-oxidation pathway were decreased. Our gene expression data in cumulus cells suggest the disruption of endoplasmic reticulum stress, apoptosis and cellular stress response pathways during IVM. Furthermore, a total of 19 miRNAs were significantly altered by the maturation process in cumulus cells. These results indicate some new negative influences of the in vitro system in cumulus-oocyte complexes, demonstrating the occurrence of functional disruption in lipid metabolism and stress pathways and showing evidences suggesting the occurrence of altered mitochondrial activity and energy metabolism during IVM, with a massive dysregulation of the corresponding transcripts in the surrounding cumulus cells.

Oleic acid (OA) and cis-9, trans-11 conjugated linoleic acid (c9t11-CLA) are fatty acids found in beef with beneficial effects in human health. This study investigated differentially abundant proteins (DAPs) in skeletal muscle of bovines with extreme values of OA, and c9t11-CLA. For each one of the fatty acids, twenty muscle samples were divided into two groups (N = 10_High; N = 10_Low) and analyzed by high definition mass spectrometry. We identified 103 and 133 DAPs between the groups for each fatty acid. We found 64 and 45 up-regulated and 39 and 68 down-regulated proteins for OA and c9t11-CLA, respectively. Comparative analysis between proteomic and transcriptomic data revealed eight and ten genes with a consistent between mRNA expression levels and protein abundance for OA and c9t11-CLA, respectively. Unconventional myosin-Id (MYO1D), mineralocorticoid receptor (NR3C2), geranylgeranyl transferase type-2 subunit-alpha (RABGGTA), and uveal autoantigen with coiled-coil domains and ankyrin repeats (UACA) were found as putative candidate proteins for OA content. Fatty acid synthase (FASN), tubulin alpha-4A chain (TUBA4A), vinculin (VCL), NADH dehydrogenase 1 alpha subcomplex 5 (NDUFA5), and prefoldin subunit 6 (PFDN6) for c9t11-CLA. Our findings contribute to a deeper understanding of the molecular mechanisms behind the regulation of the OA and c9t11-CLA content in cattle skeletal muscle. Significance Questions about the association between meat intake and disease incidence in humans has driven animal scientist to pursue a better understanding of the biological processes associated with differences in the intramuscular fat composition. The beneficial effects of oleic acid and conjugated linoleic acid in human health have been demonstrated by improving the immune system and preventing atherosclerosis, different types of cancers, hypertension, and diabetes. Previous genome-wide association and gene expression studies identified genomic regions and differentially expressed genes associated with the fatty acid profile in skeletal muscle. In this work, differences were evaluated at the protein level. The use of a label-free quantitative proteomic approach, compared with muscle transcriptome results obtained by RNA-sequencing, allowed us to earn new insights into the variability in fatty acid deposition in skeletal muscle of farm animals. This study opens new avenues to explore the effect of the fatty acids in the skeletal muscle of livestock animals, which is associated with nutritional values of the meat, and perhaps to understand the mechanisms correlated with metabolic diseases in other species.

A rapid and selective method to determine p-chloroaniline (pCA) in gel, 2 % aqueous solutions, and 0.12 % oral rinse formulas of chlorhexidine digluconate (CHX) used in dentistry treatments was developed and validated. The method was appropriate for ensuring that these products are in accordance with current legislation. Furthermore, the precipitate formed when 2 % CHX is added to sodium hypochlorite was investigated to verify whether this mixture forms pCA. To quantify pCA, liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS) was used and the following m/z 127.9/93.0 and 127.9/111.0 were used as qualifier and quantifier transitions, respectively. The LC separation, using a C18 column proved highly efficient for pCA and its isomers, i.e., m-chloroaniline and o-chloroaniline. Multiple reaction monitoring provided the proper selectivity and specificity for the method. Commercial aqueous solutions, gels, and oral rinses containing CHX were analyzed, and their pCA contents complied with those recommended by the European and United States Pharmacopeias. The method was also able to detect pCA in the precipitate and its concentration is below 0.1 %.

The pathways involved in intramuscular fat (IMF) deposition in Longissimus dorsi muscle were investigated using an integrated transcriptome-assisted label-free quantitative proteomic approach by High Definition Mass Spectrometry. We quantified 1582 proteins, of which 164 were differentially abundant proteins (DAPs, p 0.05) between animals with high (H) and low (L) genomic estimated breeding values (GEBV) for IMF content. Ingenuity pathway analysis (IPA) revealed that these DAPs were mainly involved in glycolysis metabolism, actin cytoskeleton signaling, cell-cell adherens junction and pathways for MAPK and insulin. A comparative study between transcriptomic (mRNA) and proteomic data showed 17 differentially expressed genes corresponding to DAPs, of which three genes/proteins did not agree on the direction of the fold change between groups. Moreover, we investigated microRNAs data to explain these differences in fold change direction, being able to unravel two of the three unexpected mRNA/protein relationships. Results demonstrated that changes in protein/mRNA levels of sarcomere organization, intracellular signal transduction and regulation of actin cytoskeleton, are involved in IMF deposition. These findings provide a deeper understanding of the highly complex regulatory mechanisms involved in IMF deposition in cattle and indicate target pathways for future studies.Intramuscular fat is the amount of fat deposited inside muscle and plays an important role in human health and meat quality attributes, influencing energy metabolism of skeletal muscle, as well as, tenderness, flavor, and juiciness of beef. We performed for the first time the utilization of integrated transcriptome-assisted label-free quantitative proteomic approach using High Definition Mass Spectrometry for characterization of the changes in the proteomic profile of the Longissimus dorsi muscle associated with intramuscular fat deposition in cattle. Furthermore, we compared the muscle proteome with the muscle transcriptome (mRNA and microRNAs), obtained by RNA-sequencing, to better understand the relationship between expression of mRNAs and proteins and to unravel essential biological mechanisms involved in bovine skeletal muscle IMF deposition.

Fluoroquinolones (FQs) are synthetic antimicrobials commonly used in intensive poultry farming to treat chronic respiratory disease, colibacillosis, and fowl cholera. As these drugs are also important in treating infections in humans, their use in poultry has been restricted in some countries to avoid the development of antimicrobial-resistant bacteria. In this work, a novel simple and efficient LC-MS/MS method using solid phase extraction (SPE), ESI ionization and multiple reaction mode (MRM) has been developed and validated to determine the two most common FQ residues: enrofloxacin (ENR) and ciprofloxacin (CIP) in chicken target tissues (muscle, liver, kidney, fat and skin) and in plasma. The method is shown to be sensitive, with limits of quantification (LOQ) between 1 ng g−1 and 5 ng g−1, recoveries 93–115% and correlation coefficients greater than 0.99. This novel methodology allows faster and simpler sample workflow compared to previously reported methodologies, making it ideal for high-throughput screening and confirmation of both ENR and CIP meeting the international regulation levels.

Focusing on the O2 class, a set of crude oils from Llanos Orientales Basin, Colombia, were classified in terms of biodegradation levels using negative ion mode electrospray Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) and chemometric analysis. The O2 class, which is mainly composed of naphthenic carboxylic acids, was monitored because these polar crude oil constituents are known to be substantially affected by microbial activity. Principal component analysis (PCA) applied on the O2 profile was able to classify the crude oils into three groups: biodegraded, mixture, and non-biodegraded. From the relative abundances of the O2 class, a clear trend on acid distribution could be directly correlated with biodegradation: a rising in abundance of saturated acids with low double-bond equivalent (DBE) values (despite the lowering observed for fatty acids with DBE = 1), expressed by the A/C index. The combined use of two indexes, the A/C index and a new index also based on saturated acid a...

Laboratorio de Quimica Orgânica Fina Departamento de Fisica, Quimica e Biologia Universidade Estadual Paulista (UNESP), Sao Paulo, 19060-900

Made available in DSpace on 2018-12-11T16:46:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-07-01 School of Food Engineering Department of Food Sciences University of Campinas – UNICAMP ThoMSon Mass Spectrometry Laboratory Institute of Chemistry University of Campinas - UNICAMP Department of Toxicology and Pharmacology Faculty of Veterinary Medicine Universidad Complutense de Madrid Biosciences Languages and Exact Sciences Institute – (IBILCE) Júlio de Mesquita Filho University - UNESP Biosciences Languages and Exact Sciences Institute – (IBILCE) Júlio de Mesquita Filho University - UNESP

Easy sonic-spray ionization mass spectrometry (EASI–MS) allows for direct and fast MS analysis of samples in ambient conditions with little or no sample preparation, therefore offering unprecedented simplicity, speed, and ease of use. EASI–MS has been shown to access the quality, type, and adulteration of biofuels and vegetable oils. Herein, EASI–MS is shown to quantitate and monitor the quality of soybean biodiesel/petrodiesel (Bn) blends. For adulteration, admixture of the parent oil has been tested and nearly instantaneous and direct EASI(+)–MS detection of as little as 1% (w/w) of soybean oil in biodiesel/petrodiesel blends was achieved. Linear analytical curves (r > 0.98) were also obtained for the quantitation of Bn blends, and the EASI(+)–MS quantitation results were compared to those obtained by nuclear magnetic resonance (NMR) spectroscopy and mid-infrared (IR) spectroscopy.

Seven gas oil (GO) cuts from the same atmospheric petroleum residuum were obtained by molecular distillation at final temperatures of 490.0 and 503.2 °C (medium GO), 522.5 and 549.5 °C (heavy GO), and 583.7, 622.4, and 662.2 °C (extra heavy GO). The detailed chemical composition of these samples was investigated using comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (GC × GC–TOFMS) and electrospray ionization ultrahigh-resolution and -accuracy Fourier transform ion cyclotron resonance mass spectrometry (ESI FT-ICR MS). Many compound classes were identified by GC × GC–TOFMS, such as tri-, tetra-, and pentacyclic terpanes, steranes, and secohopanes; several polycyclic aromatic hydrocarbons (PAHs), such as fluorene, phenanthrene, pyrene, and benzo[g,h,i]perylene; sulfur compounds, e.g., alkylbenzothiophenes, alkyldibenzothiophenes, and alkylbenzonaphthothiophenes; and alkylphenols. With ESI FT-ICR MS in both the positive- and negative-ion modes, many polar components tha...

Oxidation is the major cause of fat and oil deterioration which produces rancid and unpleasant flavors that drastically reduces their quality and nutritional value. The primary oxidation products are triacylglycerols (TAG) containing unsaturated fatty acyl hydroperoxides formed by a chain process occurring through free radical intermediates. The Rancimat method is currently the gold standard for measuring the oxidative stability of oils and fats. In this report, easy ambient sonic-spray ionization mass spectrometry (EASI-MS) is demonstrated to function as a direct (no pre-separation or sample preparation steps), fast and accurate method to monitor oils and fats oxidation, providing detailed overviews of the most immediate TAG hydroperoxide products. The EASI-MS results were compared to those obtained via the classical Rancimat test.

Venturi easy ambient sonic spray ionization mass spectrometry in both itsliquid(VL-EASI-MS) and solid sample modes (VS-EASI-MS) is shown to provide nearly immediate and secure typification of woods, as demonstrated for Mahogany, an endangered and mostvaluabletypeoftropical wood. Thisreddishwood displaysuniquephytochemical markers(phragmalin-typelimonoids) which are rapidly detected from the wood surface by VS-EASI-MS or from a simple methanol extract of a tiny wood chip by VL-EASI-MS. Unique profiles were obtained for Mahogany (Swietenia macrophylla) whereas genuine samples of six other similar types of woods, which are commonly falsified by artificial coloring and commercialized as Mahogany, display also typical but dissimilar pythochemical profiles as compared to that of the authentic wood. Variable and atypical chemical profiles were observed for artificially colored woods. Secure chemical characterization via VS-EASI-MS or Vs-EASI-MS fingerprints of Mahogany and other types of woods with similar appearance should help to control the illegal logging and trade of this and other endangered woods and their falsification, and to create certified standards. Copyright © 2012 John Wiley & Sons, Ltd.

Easy ambient sonic-spray ionization mass spectrometry (EASI–MS) was used to follow the maturation of Jatropha curcas L. seeds via the monitoring of the triacylglycerides (TAG) profile of the oil. Results show that TAG composition is significantly modified during seed development but remains nearly unchanged during storage. The EASI–MS oil analysis performed herein is simple, requires just a tiny droplet of the oil and is performed without any pre-separation or chemical manipulation. The oil from Jatropha gossypifolia L. was also evaluated, and a very different TAG profile was obtained.

The production of structured lipids (SLs) by the acidolysis of soybean oil (SO) with a free fatty acid (FFA) mixture obtained from Brazilian sardine oil, catalysed by Rhizomucor miehei lipase (Lipozyme RM IM) in a solvent-free medium, was optimised by response surface methodology (RSM) using a three-factor central composite rotatable design. The best reaction conditions to achieve an adequate n-6/n-3 FA ratio were: sardine-FFA:SO mole ratio of 3:1, initial water content of the enzyme of 0.87% w/w, reaction time of 12 h, reaction temperature of 40 °C and 10% by weight of the enzyme (% w/w). Under these conditions, the incorporation of eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) into the soybean oil reached 9.2% (% of the total FAs), leading to a significant reduction in the n-6/n-3 FA ratio from 11:1 to 3:1. Analysis of variance (ANOVA) showed that 95% (R2 = 0.95) of the observed variation was explained by the model. Lack of fit analysis revealed a non-significant value for the model equation, indicating that the regression equation was adequate for predicting the degree of EPA + DHA incorporation under any combination of values of the variables. Easy ambient sonic-spray ionisation mass spectrometry (EASI-MS) was used for instantaneous characterisation of TAGs. After the enzymatic reaction, a great variety of new TAGs were formed containing EPA, DHA or both in the same molecule.

Biodiesel is composed of saturated and unsaturated methyl esters of fatty acids formed via transesterification of vegetable oil or animal fat. This chemical nature makes biodiesel much more susceptible to oxidation or autoxidation during long-term storage than conventional petrodiesel. Increasing resistance to oxidation therefore requires the use of antioxidants. This paper investigates the effectiveness of the N,N′-di-sec-butyl-p-phenylenediamine (1) as a antioxidant to improve the oxidative stability of soybean, sunflower and canola biodiesel. Results indicate that the induction period for oxidation of soybean and canola biodiesel is significantly improved with the addition of only 0.2 ppm of the 1, reaching the oxidative stability specification (6 h by the Rancimat test). For sunflower biodiesel, 2.0 ppm of the 1 was necessary. This additive is also shown to be efficiently detected by the easy ambient sonic-spray ionization mass spectrometry technique, indicating that 1 and its analogues and derivative...

Mass spectrometry has recently undergone a second contemporary revolution with the introduction of a new group of desorption/ionization (DI) techniques known collectively as ambient mass spectrometry. Performed in an open atmosphere directly on samples in their natural environments or matrices, or by using auxiliary surfaces, ambient mass spectrometry (MS) has greatly simplified and increased the speed of MS analysis. Since its debut in 2004 there has been explosive growth in the applications and variants of ambient MS, and a very comprehensive set of techniques based on different desorption and ionization mechanisms is now available. Most types of molecules with a large range of masses and polarities can be ionized with great ease and simplicity with the outstanding combination of the speed, selectivity, and sensitivity of MS detection. This review describes and compares the basis of ionization and the concepts of the most promising ambient MS techniques known to date and illustrates, via typical analytical and bioanalytical applications, how ambient MS is helping to bring MS analysis deeper than ever into the "real world" open atmosphere environment--to wherever MS is needed.

Potentially bioactive N-(aminoalkyl)lactamic amino acids and esters were synthesized in satisfactory to good yields by S(N)Ar reactions of aromatic acids with N-(3-aminopropyl)lactams followed by esterification with tertiary amino alcohols. The addition-elimination S(N)Ar mechanism was confirmed by NMR and MS measurements.

Made available in DSpace on 2015-10-21T13:10:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-05-26. Added 1 bitstream(s) on 2015-10-22T09:53:04Z : No. of bitstreams: 1 WOS000355183900034.pdf: 1411034 bytes, checksum: c0a385ed9382e3e64549cc87bc043782 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Golden retriever muscular dystrophy (GRMD) provides the best animal model for characterizing the disease progress of the human disorder, Duchenne muscular dystrophy (DMD). The purpose of this study was to determine steroid hormone concentration profiles in healthy golden retriever dogs (control group - CtGR) versus GRMD-gene carrier (CaGR) and affected female dogs (AfCR). Therefore, a sensitive and specific analytical method was developed and validated to determine the estradiol, progesterone, cortisol, and testosterone levels in the canine serum by isotope dilution liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). To more accurately understand the dynamic nature of the serum steroid profile, the fluctuating levels of these four steroid hormones over the estrous cycle were compared across the three experimental groups using a multivariate statistical analysis. The concentration profiles of estradiol, cortisol, progesterone, and testosterone revealed a characteristic pattern for each studied group at each specific estrous phase. Additionally, several important changes in the serum concentrations of cortisol and estradiol in the CaGR and AfCR groups seem to be correlated with the status and progression of the muscular dystrophy. A comprehensive and quantitative monitoring of steroid profiles throughout the estrous cycle of normal and GRMD dogs were achieved. Significant differences in these profiles were observed between GRMD and healthy animals, most notably for estradiol. These findings contribute to a better understanding of both dog reproduction and the muscular dystrophy pathology. Our data open new venues for hormonal behavior studies in dystrophinopathies and that may affect the quality of life of DMD patients. Univ Campinas UNICAMP, Inst Chem, ThoMSon Mass Spectrometry Lab, Campinas, SP, Brazil AB SCIEX Brazil, Sao Paulo, SP, Brazil Sao Paulo State Univ UNESP, Fac Vet Med, DAPSA, Aracatuba, SP, Brazil Univ Sao Paulo, FZEA, Pirassununga, SP, Brazil Univ Sao Paulo, Fac Vet Med &Anim Sci FMVZ, Sao Paulo, SP, Brazil Purdue Univ, Dept Chem, W Lafayette, IN 47907 USA Sao Paulo State Univ UNESP, Fac Vet Med, DAPSA, Aracatuba, SP, Brazil FAPESP: 2007/51222-2 FAPESP: 2010/15167-0 FAPESP: 2010/51677-2

The plasma levels of corticosteroids and sex steroids during pregnancy are key indicators of mammalian placental function and the onset of parturition. Steroid hormones are believed to be disturbed in pregnancies produced using assisted reproductive technologies (ARTs) due to placental dysfunction and the frequently observed lack of parturition signals. To elucidate the plasma steroid dynamics, a liquid chromatography-tandem mass spectrometry method was developed and used to determine the levels of corticosteroids (corticosterone, 11-deoxycortisol, and cortisol) and their direct precursors (progesterone and 17α-OH-progesterone) as well as sex steroids (androstenedione, estrone, estrone sulfate, testosterone, and 17β-estradiol) in bovine plasma. The levels of these 10 steroids in recipient cows carrying naturally conceived (control), in vitro fertilized (IVF), or cloned (somatic cell nuclear transfer) conceptuses were compared during late-term pregnancy (30 days before parturition), during near-term pregnancy (1 day before parturition), and on the day of parturition (day 0). Significant differences were observed among the corticosteroid levels: higher levels of corticosterone, 11-deoxycortisol, and cortisol were detected in cloned pregnancies at day 30; lower levels of corticosterone were observed in ART-derived pregnancies at days 1 and 0; and estrone and estradiol levels were higher in IVF pregnancies throughout the final development. These results suggested an upregulation of the P450C11 and P450C21 enzymes 30 days before parturition in somatic cell nuclear transfer pregnancies and an overactivation of the aromatase enzyme in IVF pregnancies. Taken together, the monitoring of multiple steroid hormones revealed that the pregnancies obtained using ART exhibited plasma steroid concentration dynamics compatible with the dysregulation of steroidogenic tissues.

The introduction of precise analytical technologies in the biological area is needed to increase the knowledge of the fundamental processes occurring in the animal and human domain. The objective of this study was to develop a highly sensitive and selective method based on liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) using electrospray ionization for the simultaneous detection and quantification of melatonin (MEL) and its metabolites, 6-hydroxymelatonin (6-HMEL), N2-acetyl-N1-formyl-5-methoxykynuramine (AFMK) and N1-acetyl-5-metoxykynuramine (AMK). Using this methodology, we have studied the role of melatonin and its metabolites in the mammalian embryo in vitro production system, which is fundamental due to the antioxidant and signaling roles of these compounds. Samples comprised of bovine follicular fluid (FF) and tissue culture medium (TCM 199). The optimized procedure uses liquid–liquid extraction with MS monitoring via selective reaction monitoring (SRM). Low limits of detection/quantification were obtained, 3/10 pg mL−1 for MEL, AFMK and AMK and 30/100 pg mL−1 for 6-HMEL, respectively using deuterated melatonin (MEL-d4) as the internal standard (IS). Validation and correlation coefficients were higher than 0.999 and recoveries were 80–108%. Precision was evaluated as repeatability and intermediate precision with relative standard deviation values

Brazil nut oil is one of the important Amazonian natural products in the global market. Despite its health benefits and applications in food and cosmetic industries, authentication and quality control of the oil are far from satisfactory. Several samples of Amazonian Brazil nut oil (authentic oils of different geographic origins, commercial oils, and oils adulterated with soybean oil) were evaluated by easy ambient sonic-spray ionization mass spectrometry (EASI-MS). The samples were characterized on the basis of triacylglycerol profiles, and their major ions were subjected to chemometric treatment (principal component analysis). The ambient mass spectrometry analysis and the set of major ions allowed the characterization of authentic oils, commercial oils, and adulterated oils (5% soybean oil) in a few minutes and without sample preparation. Therefore, it is shown that quality control of Amazonian vegetable oils can be readily accomplished by EASI-MS.

Venturi easy ambient sonic spray ionization mass spectrometry in both its liquid (V(L) -EASI-MS) and solid sample modes (V(S) -EASI-MS) is shown to provide nearly immediate and secure typification of woods, as demonstrated for Mahogany, an endangered and most valuable type of tropical wood. This reddish wood displays unique phytochemical markers (phragmalin-type limonoids) which are rapidly detected from the wood surface by V(S) -EASI-MS or from a simple methanol extract of a tiny wood chip by V(L) -EASI-MS. Unique profiles were obtained for Mahogany (Swietenia macrophylla) whereas genuine samples of six other similar types of woods, which are commonly falsified by artificial coloring and commercialized as Mahogany, display also typical but dissimilar pythochemical profiles as compared to that of the authentic wood. Variable and atypical chemical profiles were observed for artificially colored woods. Secure chemical characterization via V(S) -EASI-MS or V(s) -EASI-MS fingerprints of Mahogany and other types of woods with similar appearance should help to control the illegal logging and trade of this and other endangered woods and their falsification, and to create certified standards.

An ambient ionization/desorption technique, namely, easy ambient sonic-spray ionization mass spectrometry (EASI), has been applied to crude oil samples. From a single droplet of the sample placed on an inert surface, EASI(+/-) is shown to promote efficient desorption and ionization of a myriad of polar components via the action of its cloud of very minute supersonic bipolar charged droplets. The gaseous [M + H](+) and [M - H](-) ions concurrently formed by EASI(+/-) were analyzed by Fourier transform mass spectrometry (FT-ICR MS), and a total of approximately 6000 acidic and basic components have been attributed. EASI(+/-) FT-ICR MS of crude oils is show to be almost as fast as ESI(+)/ESI(-) FT-ICR MS, providing similar compositional information of polar components and spectral quality comparable to that of a commercial nonochip-based robotic ESI device. EASI(+/-) requires no sample workup thus eliminating risks of contamination during sample manipulation and memory effects because of carry over in pumping ESI lines. More importantly, EASI(+/-) is a voltage-free ionization technique therefore eliminating risks of redox processes or duality of ionization mechanisms that can be observed in voltage-assisted processes. Data visualization via typical petroleomic plots confirms the similarity of the compositional information provided by EASI(+/-) compared to ESI(+)/ESI(-). The ambient EASI(+/-) FT-ICR MS method requires no voltage switching in changing the ion polarity mode, offering a workup, heating and voltage-free protocol for petroleomic studies performed at open atmosphere directly on the undisturbed crude oil sample.

Although the production of patulin in apple fruits is mainly by Penicillium expansum, there is no information on the ability of heat resistant moulds that may survive pasteurization to produce this mycotoxin in juice packages during storage and distribution. In this study, the production of patulin by Byssochlamys spp (Byssochlamys nivea FRR 4421, B. nivea ATCC 24008 and Byssochlamys fulva IOC 4518) in cloudy and clarified apple juices packaged in laminated paperboard packages or in polyethylene terephthalate bottles (PET) and stored at both 21 degrees C and 30 degrees C, was investigated. The three Byssochlamys strains were able to produce patulin in both cloudy and clarified apple juices. Overall, the lower the storage temperature, the lower the patulin levels and mycelium dry weight in the apple juices (p0.05). The greatest variations in pH and degrees Brix were observed in the juices from which the greatest mycelium dry weights were recovered. The maximum levels of patulin recovered from the juices were ca. 150 microg/kg at 21 degrees C and 220 microg/kg at 30 degrees C. HPLC-UV, HPCL-DAD and mass spectrometry analyses confirmed the ability of B. fulva IOC 4518 to produce patulin. Due to the heat resistance of B. nivea and B. fulva and their ability to produce patulin either in PET bottles or in laminated paperboard packages, the control of contamination and the incidence of these fungi should be a matter of concern for food safety. Control measures taken by juice industries must also focus on controlling the ascospores of heat resistant moulds.

The oil from the seeds of the “Andiroba” tree, which is found throughout most tropical America, contains high levels of unsaturated triacylglycerols (TAG), which makes it attractive to the cosmetics industry. A significant amount of limonoids also confers to this Amazonian oil several pharmaceutical and medical properties. In addition, the oil is also a potential feedstock for biodiesel production, and its many uses have intensified its extractive exploitation in recent years. Herein we report on the characterization of the TAG, free fatty acids (FFA) and limonoid profiles of the Andiroba oil via mass spectrometry (MS) fingerprinting using direct electrospray ionization mass spectrometry (ESI-MS). An ambient desorption/ionization technique known as easy ambient sonic-spray ionization (EASI-MS) was also evaluated with similar results. ESI-MS was performed either for a methanolic solution of a few microliters of the fresh oil or from a simple aqueous extract whereas EASI-MS was applied directly to a droplet of the oil resting on a paper surface. The efficacy of these MS fingerprinting techniques requiring no pre-separation and no or very simple sample preparation protocols was investigated and compared for the typification and quality of this valuable Amazonian oil.

This innovative mass spectrometric technique provides simple, quick, and reliable quantitative analysis of triacylglycerols in vegetable oils and fats. The easy ambient sonic-spray ionization mass spectrometry (EASI-MS) technique provides simple, quick, and reliable qualitative analysis of triacylglycerols (TAG) in vegetable oils. The use of this ambient mass spectrometry technique, which does not require separation or derivatization before analysis, for precise quantitative analyses has yet to be demonstrated. In this study, the TAG composition of edible vegetable oils, hydrogenated vegetable oils and cocoa butter was quantitated by EASI-MS. The results were compared with those obtained by theoretical predictions of the TAG composition performed by a software projection, which uses a mathematical algorithm of distribution of the fatty acids (FA) in the TAG molecule, and by the direct gas chromatography method (GC-FID). Acceptable correlation coefficients were observed between the three methods during the analysis of vegetable oils and hydrogenated vegetable oils. EASI-MS offers not only simplicity, speed and selectivity for oil analysis but also precise quantitation of the TAG composition. Therefore, EASI-MS is a promising substitute for the more demanding and time-consuming standard protocols, which require separation and sometimes derivatization before analysis.

Quantitation of progesterone (P(4)) in biological fluids is often performed by radioimmunoassay (RIA), whereas liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) has been used much less often. Due to its autoconfirmatory nature, LC-MS/MS greatly minimizes false positives and interference. Herein we report and compare with RIA an optimized LC-MS/MS method for rapid, efficient, and cost-effective quantitation of P(4) in plasma of cattle with no sample derivatization. The quantitation of plasma P(4) released from three nonbiodegradable, commercial, intravaginal P(4)-releasing devices (IPRD) over 192 h in six ovariectomized cows was compared in a pairwise study as a test case. Both techniques showed similar P(4) kinetics (P > 0.05) whereas results of P(4) quantitation by RIA were consistently higher compared with LC-MS/MS (P < 0.05) due to interference and matrix effects. The LC-MS/MS method was validated according to the recommended analytical standards and displayed P(4) limits of detection (LOD) and quantitation (LOQ) of 0.08 and a 0.25 ng/mL, respectively. The high selective LC-MS/MS method proposed herein for P(4) quantitation eliminates the risks associated with radioactive handling; it also requires no sample derivatization, which is a common requirement for LC-MS/MS quantitation of steroid hormones. Its application to multisteroid assays is also viable, and it is envisaged that it may provide a gold standard technique for hormone quantitation in animal reproductive science studies.

A fast and reliable method is presented for the analysis of vegetable oils. Easy ambient sonic-spray ionization mass spectrometry (EASI-MS) is shown to efficiently desorb and ionize the main oil constituents from an inert surface under ambient conditions and to provide comprehensive triacylglyceride (TAG) and free fatty acid (FFA) profiles detected mainly as either [TAG + Na](+) or [FFA-H](-) ions. EASI(+/-)-MS analysis is simple, easily implemented, requires just a tiny droplet of the oil and is performed without any pre-separation or chemical manipulation. It also causes no fragmentation of TAG ions hence diacylglyceride (DAG) and monoacylglyceride (MAG) profiles and contents can also be measured. The EASI(+/-)-MS profiles of TAG and FFA permit authentication and quality control and can be used, for instance, to access levels of adulteration, acidity, oxidation or hydrolysis of vegetable oils in general.