Chemical Ionization Tandem Mass Spectrometry Research Articles

Omega-3 Long-Chain Polyunsaturated Fatty Acids in Nonseafood and Estimated Intake in the USA: Quantitative Analysis by Covalent Adduct Chemical Ionization Mass Spectrometry.

Omega-3 long-chain polyunsaturated fatty acids (LCPUFA) play critical roles in human development and health. Their intake is often effectively estimated solely based on seafood consumption, though the high intake of terrestrial animal-based foods with minor amounts of LCPUFA may be significant. Covalent adduct chemical ionization (CACI) tandem mass spectrometry is one approach for de novo structural and quantitative analysis of minor unsaturated fatty acids (FA), for which standards are unavailable. Here, CACI-MS and MS/MS are used to identify and quantify minor omega-3 LCPUFA of terrestrial animal foods based on the application of measured response factors (RFs) to various FA. American mean intakes of pork, beef, chicken, and eggs contribute 20, 27, 45, and 71 mg/day of docosahexaenoic acid (DHA), respectively. The estimated intake of omega-3 DHA, eicosapentaenoic acid, and docosapentaenoic acid from nonseafood sources is significant, at 164, 103, and 330 mg/day, greater than most existing estimates of omega-3 LCPUFA intake.

Determination of Patulin in Apple Juice and Apple-Derived Products Using a Robotic Sample Preparation System and LC-APCI-MS/MS.

Patulin, a toxic mycotoxin, can contaminate apple-derived products. The FDA has established an action level of 50 ppb (ng/g) for patulin in apple juice and apple juice products. To effectively monitor this mycotoxin, there is a need for adequate analytical methods that can reliably and efficiently determine patulin levels. In this work, we developed an automated sample preparation workflow followed by liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry (LC-APCI-MS/MS) detection to identify and quantify patulin in a single method, further expanding testing capabilities for monitoring patulin in foods compared to traditional optical methods. Using a robotic sample preparation system, apple juice, apple cider, apple puree, apple-based baby food, applesauce, fruit rolls, and fruit jam were fortified with 13C-patulin and extracted using dichloromethane (DCM) without human intervention, followed by an LC-APCI-MS/MS analysis in negative ionization mode. The method achieved a limit of quantification of 4.0 ng/g and linearity ranging from 2 to 1000 ng/mL (r2 > 0.99). Quantitation was performed with isotope dilution using 13C-patulin as an internal standard and solvent calibration standards. Average recoveries (relative standard deviations, RSD%) in seven spike matrices were 95% (9%) at 10 ng/g, 110% (5%) at 50 ng/g, 101% (7%) at 200 ng/g, and 104% (4%) at 1000 ng/g (n = 28). The ranges of within-matrix and between-matrix variability (RSD) were 3-8% and 4-9%, respectively. In incurred samples, the identity of patulin was further confirmed with a comparison of the information-dependent acquisition-enhanced product ion (IDA-EPI) MS/MS spectra to a reference standard. The metrological traceability of the patulin measurements in an incurred apple cider (21.1 ± 8.0 µg/g) and apple juice concentrate (56.6 ± 15.6 µg/g) was established using a certified reference material and calibration data to demonstrate data confidence intervals (k = 2, 95% confidence interval).

Differentiating the aromatic positional isomers of methylbuphedrones and methoxybuphedrones via chemical ionization-mass spectrometry.

Discrimination of aromatic positional isomers of methylbuphedrones and methoxybuphedrones was successfully achieved. Meta isomers were discriminated by chemical ionization-tandem mass spectrometry (CI-MS/MS) using acetonitrile as a reagent gas. Furthermore, all the aromatic positional isomers were discriminated by CI-MS/MS using vinyltrimethylsilane as a reagent gas.

PM2.5-Bound Organophosphate Flame Retardants in Hong Kong: Occurrence, Origins, and Source-Specific Health Risks.

Organophosphate flame retardants (OPFRs) are emerging organic pollutants in PM2.5, which have caused significant public health concerns in recent years, given their potential carcinogenic and neurotoxic effects. However, studies on the sources, occurrence, and health risk assessment of PM2.5-bound OPFRs in Hong Kong are lacking. To address this knowledge gap, we characterized 13 OPFRs in one-year PM2.5 samples using gas chromatography-atmospheric pressure chemical ionization tandem mass spectrometry. Our findings showed that OPFRs were present at a median concentration of 4978 pg m-3 (ranging from 1924 to 8481 pg m-3), with chlorinated OPFRs dominating and accounting for 82.7% of the total OPFRs. Using characteristic source markers and positive matrix factorization, we identified one secondary formation and five primary sources of OPFRs. Over 94.0% of PM2.5-bound OPFRs in Hong Kong were primarily emitted, with plastic processing and waste disposal being the leading source (61.0%), followed by marine vessels (14.1%). The contributions of these two sources to OPFRs were more pronounced on days influenced by local pollution emissions (91.9%) than on days affected by regional pollution (44.2%). Our assessment of health risks associated with human exposure to PM2.5-bound OPFRs indicated a low-risk level. However, further source-specific health risk assessment revealed relatively high noncarcinogenic and carcinogenic risks from chlorinated OPFRs emitted from plastic processing and waste disposal, suggesting a need for more stringent emission control of OPFRs from these sources in Hong Kong.

An Analytical Method for Determining N-Nitrosodimethylamine and N-Nitrosodiethylamine Contamination in Irbesartan, Olmesartan and Metformin by UPLC-APCI-MS/MS in Tablet Dosage Form.

N-nitrosamine pollutants are probable carcinogens. Regulatory agencies declared their presence in the drugs unsafe for human consumption and demanded their recall. Using ultra-performance liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry (UPLC-APCI-MS/MS) in tablet dosage form based on International Conference on Harmonization (ICH) tripartite guideline criteria, we aim to develop and test a new approach for identifying and validating nitrosamine-contaminants, N-nitrosodimethylamine (NDMA) and N-nitrosodiethylamine (NDEA) in irbesartan, olmesartan and metformin. The column was Phenomenex Luna-C18, 100 × 3.0mm and 3.0μm. A mobile gradient phase of formic acid in either water or methanol separated the impurities. NDMA and NDEA had retention times of 0.85 and 2.55min, respectively. The detector's linearity was established at concentrations ranging from 0.6 to 100ng/mL. R2 for NDMA and NDEA were 0.9996 and 0.9998, respectively, with a linear response function established at 0.6-100ng/mL. Limit of detection and limit of quantification for NDMA and NDEA were 0.35, 0.29 and 0.55, 0.37ng/mL, respectively. On average, recovery rates for NDMA and NDEA ranged from 96.0 to 98.4 and 96.2 to 98.0%, respectively. The relative standard deviation for NDMA and NDEA was 3.46 and 2.69, respectively. According to the ICH guidelines, the developed method was quick, sensitive and valid. The pharmaceutical formulations of irbesartan, olmesartan and metformin may be regularly examined using the approach provided here.

Determination of eight herbicide residues in foodstuffs of plant origin by gas chromatography–atmospheric pressure chemical ionization–tandem mass spectrometry

This study uses gas chromatography–atmospheric pressure chemical ionization–tandem mass spectrometry (GC–APCI–MS/MS) to detect and identify 8 herbicide residues in foodstuffs of plant origin. In contrast to the extensive fragmentation caused by traditional electron impact ionization (EI) with high energy, APCI is a soft ionization technique. Therefore, more molecular ions and/or protonated molecules are present, improving the sensitivity and selectivity of analytical methods. The standard calibration curves ranged from 0.5 to 50 ng/mL; recoveries were in the range of 76.3–108.5%. The limit of quantification was 1.0 µg/kg for all samples. The results demonstrate the potential of GC–APCI–MS/MS to routinely detect and identify 8 herbicide residues in foodstuffs of plant origin.

Analysis of N-nitrosamines and N-nitrosatable substances from baby bottle rubber teats by liquid chromatography tandem mass spectrometry

A simple and sensitive method based on liquid chromatography-atmospheric pressure chemical ionisation-tandem mass spectrometry (LC-APCI-MS/MS) was developed and validated to determine the levels of 13N-nitrosamines and N-nitrosatable substances migrated from rubber teats into artificial saliva. The migration test from rubber teats was conducted at 40 °C and for 24 h in artificial saliva, and the migrated artificial saliva solution was analysed by liquid chromatography tandem mass spectrometry (LC-MS/MS) without further extracting steps. The sensitivity of N-nitrosamines was examined by applying atmospheric chemical ionisation and electrospray ionisation to optimise the mass spectrometric conditions, and the atmospheric chemical ionisation (APCI) mode exhibited 1.6–19 times higher sensitivity. Method validation showed acceptable linearity, precision, and accuracy, and the detection and quantification limits were 0.07–0.35 and 0.24–1.1 μg kg−1, respectively. The developed liquid chromatography-atmospheric chemical ionisation-tandem mass spectrometry method was applied to 39 domestic and imported rubber teats. From 39 samples, N-nitrosamines [N-nitrosodimethylamine (NDMA), N-nitrosomorpholine (NMOR), and N-nitroso n-methyl N-phenylamine (NMPhA)] were detected in 30 samples, with N-nitrosatable substances in 17 samples give rise to NDMA, NMOR, and N-nitrosodiethylamine. However, the levels were below the specific migration limit of Korean Standards and Specifications for Food Containers, Utensils, and Packages and EC Directive 93/11/EEC.

Development of Quantitative Chemical Ionization Using Gas Chromatography/Mass Spectrometry and Gas Chromatography/Tandem Mass Spectrometry for Ambient Nitro- and Oxy-PAHs and Its Applications

The concentration of polycyclic aromatic hydrocarbons (PAHs) in the atmosphere has been continually monitored since their toxicity became known, whereas nitro-PAHs (NPAHs) and oxy-PAHs (OPAHs), which are derivatives of PAHs by primary emissions or secondary formations in the atmosphere, have gained attention more recently. In this study, a method for the quantification of 18 NPAH and OPAH congeners in the atmosphere based on combined applications of gas chromatography coupled with chemical ionization mass spectrometry is presented. A high sensitivity and selectivity for the quantification of individual NPAH and OPAH congeners without sample preparations from the extract of aerosol samples were achieved using negative chemical ionization (NCI/MS) or positive chemical ionization tandem mass spectrometry (PCI-MS/MS). This analytical method was validated and applied to the aerosol samples collected from three regions in Northeast Asia-namely, Noto, Seoul, and Ulaanbaatar-from 15 December 2020 to 17 January 2021. The ranges of the method detection limits (MDLs) of the NPAHs and OPAHs for the analytical method were from 0.272 to 3.494 pg/m3 and 0.977 to 13.345 pg/m3, respectively. Among the three regions, Ulaanbaatar had the highest total mean concentration of NPAHs and OPAHs at 313.803 ± 176.349 ng/m3. The contribution of individual NPAHs and OPAHs in the total concentration differed according to the regional emission characteristics. As a result of the aerosol samples when the developed method was applied, the concentrations of NPAHs and OPAHs were quantified in the ranges of 0.016~3.659 ng/m3 and 0.002~201.704 ng/m3, respectively. It was concluded that the method could be utilized for the quantification of NPAHs and OPAHs over a wide concentration range.

Method for trace determination of N-nitrosamines impurities in metronidazole benzoate using high-performance liquid chromatography coupled with atmospheric-pressure chemical ionization tandem mass spectrometry.

Genotoxic impurity control has been a great concern in the pharmaceutical industry since the recall of the large round of sartans worldwide in 2018. In these sartans, N-nitrosamines were the main contaminants in active pharmaceutical ingredients and formulations. Numerous analytical methods have been developed to detect N-nitrosamines in food, drugs, and environmental samples. In this study, a sensitive method is developed for the trace determination of N-nitrosamine impurities in metronidazole benzoate pharmaceuticals using high-performance liquid chromatography/atmospheric-pressure chemical ionization tandem mass spectrometry in the multiple reaction monitoring mode. The method was validated regarding system suitability, selectivity, linearity, accuracy, precision, sensitivity, solution stability, and robustness. The method showed good linearity with R2 ≥ 0.999 and FMandel <Ftab(95%) ranging from 0.33 to 8.00ng/ml. The low limits of detection of N-nitrosamines were in the range of 0.22-0.80ng/ml (0.0014-0.0050ppm). The low limits of quantification were in the range of 0.33-1.20ng/ml (0.0021-0.0075ppm), which were lower than the acceptable limits in metronidazole benzoate pharmaceuticals and indicated the high sensitivity of the method. The recoveries of N-nitrosamines ranged from 84% to 97%. Thus, this method exhibits good selectivity, sensitivity, and accuracy. Moreover, it is a simple, convenient, and scientific strategy for detecting N-nitrosamine impurities in pharmaceuticals to support the development of the pharmaceutical industry.

Simultaneous determination of three components of sodium nitrophenolate in foodstuffs of animal origin by high performance liquid chromatography-tandem mass spectrometry using atmospheric pressure chemical ionization

Sodium nitrophenolate (SNP) is a widely used universal growth regulator consisting of 5-nitroguaiacol sodium (5NG), 4-nitrophenol sodium (PNP), and 2-nitrophenol sodium (ONP). SNP has a positive influence on plants and animals as a feed additive that accelerates growth but is potentially hazardous to humans. SNP has been reported to be cytotoxic and mutagenic, which may increase the risk of cancer and pose a great threat to food safety. There are neither mature detection nor standard methods for the trace analysis of SNP in animal food. Therefore, the development of an accurate and precise analytical method is imperative. This innovative method has theoretical and practical significance for the control of SNP residues, offering advantages such as cost-effectiveness and time efficiency. It will be beneficial for the establishment of detection standards and management measures in foodstuffs of animal origin.In this study, a reliable method for the simultaneous determination of SNP residues in animal food (porcine muscle, chicken tissue, fish, and liver) was developed. For realizing the perfect limit of quantification, the application of back extraction coupled with high performance liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry (HPLC-APCI-MS/MS) was proposed to combine high sensitivity and high selectivity. The optimal method was as follows. First, 2.0 g samples were extracted with 10 mL 0.5 mol/L sodium hydroxide solution, followed by adjustment of the pH to acidity with 3 mol/L hydrochloric acid and the addition of sodium chloride (5.0 g) to saturate the inorganic phase. After back-extraction twice with 16 mL acetonitrile, the solution was merged and again saturated with 5 mL of sodium chloride solution. Second, the merged organic phase was cleaned up with 10 mL of n-hexane for defatting. The middle acetonitrile layer was then concentrated to nearly 1.5 mL at 40 ℃ in a N2 stream before dilution with the mobile phase to a volume of 3.0 mL and filtered. Finally, the analytes were separated on a C18 column (100 mm×4.6 mm, 3 μm) and subjected to gradient elution with a mixed solution of methanol and water. Mass spectrometric analysis, which was quantified using the external standard method, was carried out with an atmospheric pressure chemical ionization negative ion source and based on multiple reaction monitoring (MRM) mode. The key parameters, such as the extraction solvent, extraction steps, and purification method, were optimized.The calibration curves were linear in the ranges of 0.5-10 (5NG), 1.0-20 (PNP), and 2.5-50 μg/L (ONP) with correlation coefficients greater than 0.9999. The limit of quantification (LOQ) for 5NG was 1.0 μg/kg, double for PNP, and five times for ONP. The recoveries of the three different concentration levels in all the four matrices were in the range of 81.5%-98.4%, 81.5%-102%, and 81.4%-95.1%. The repeatability, expressed as the relative standard deviations (RSDs) of the three compounds, ranged from 1.51% to 5.98%, 1.10% to 8.85% and 0.91% to 8.61% (n=6). The developed method is characterized by an excellent purification effect, sensitivity, and accuracy. This method is suitable for the simultaneous and quantitative determination of SNP residues in foodstuffs of animal origin.

Determination of vitamin K encapsulated into lipid nanocarriers by dispersive liquid-liquid microextraction combined with liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry.

Dispersive liquid-liquid microextraction was used in conjunction with liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry to quantitate vitamins K1 and K2 in vitamin-fortified emulsions, and vital microextraction parameters were optimized using response surface methodology coupled with Box-Behnken design. Under optimal microextraction conditions, highly linear (R 2 > .999) calibration curves were obtained for both vitamins in a broad concentration range (1-1000 μg/L), and vitamin recoveries exceeded 90%. The detection and quantitation limits equaled 1.89 and 5.72 μg/L for vitamin K1, respectively, and 5.00 and 15.15 μg/L for vitamin K2, respectively. When applied to vitamin-K-loaded nanoemulsions and solid lipid nanoparticles, the developed method achieved excellent results, outperforming the currently employed Korean Food Code method, and therefore holding great promise for the quantitation of vitamin K in vitamin-fortified food products.

Time Course Of Dermal Anthracene Absorption Utilizing Intradermal Microdialysis

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants encountered through daily exposures, including smoking, vehicular exhaust, industry sources, and occupational settings, such as firefighting. Inhalation has received greatest attention as a major exposure route, with limited data regarding dermal absorption. PURPOSE: Utilizing the non-carcinogenic PAH, anthracene (ANT), we aimed to assess 1) time course of dermal ANT absorption and 2) effects of local skin temperature on the magnitude of dermal absorption. METHODS: Two intradermal microdialysis (MD) fibers were inserted in the ventral forearm of 6 healthy participants (32 ± 5 yrs, 5 male, 1 female). MD fibers were perfused with 10% 2-hydroxypropyl-β-cyclodextrin with lactated Ringer’s at 1 μl/min. A 2% ANT solution was applied to skin over each MD site with overlying local heaters (LH) housing laser Doppler flowmeters for assessment of skin blood flow (SkBf). LH were clamped at 33 °C during baseline dialysate sampling. Following baseline, LH were set to 1) 43 °C (Hot, HT) and 2) 33 °C as thermoneutral (TN) for the duration of the protocol. Dialysate samples were collected intermittently over 4 hours and 15 minutes, and SkBf, blood pressure and HR were recorded throughout the protocol. Atmospheric pressure chemical ionization tandem mass spectrometry was used to quantify ANT dialysate concentrations. RESULTS: Preliminary data indicate that no ANT was detected in any baseline samples. From 1 h30 to 1 h 45 min, ANT was detected in 3 of 6 and 0 of 6 samples for the HT and TN sites, respectively. One HT sample was quantifiable at 317.5 ppb. Sampling from 4 h to 4 h 15 min, ANT was detected in all samples at the HT site and quantified in one (344.9 ppb). ANT was detected in only 1 of 6 samples at the TN site during this sampling period. SkBf was significantly higher at HT versus TN at both 1 h 45 min (8.7 ± 5.7 and 29.2 ± 20.5 CVC%max, P < 0.05) and 4 h 15 min (12.8 ± 8.3 and 42.8 ± 22.3 CVC%max, P < 0.05). CONCLUSIONS: Preliminary data indicate that dermal absorption and recovery of the PAH anthracene is increased when skin is heated versus thermoneutral, increasing over time. This has potential implications for dermal absorption of contaminants in individuals working in the heat. These data also suggest that microdialysis may be an effective method of assessing dermal absorption of PAHs.

Determination and enantioselective separation of zilpaterol in human urine after mimicking consumption of contaminated meat using high-performance liquid chromatography with tandem mass spectrometry techniques.

The synthetic β-adrenoreceptor agonist zilpaterol is legitimately used as an animal feed supplement in selected countries due to its known effects on lipolysis and protein biosynthesis. These pharmacological characteristics of zilpaterol have contributed to its classification as doping agent in sport by the World Anti-Doping Agency. However, the use as a feed supplement can lead to residues of the drug in edible tissues and, possibly, also in the urine of consumers. To provide urinary elimination profiles of microdosed zilpaterol and to determine whether the ingestion of zilpaterol below or at the acceptable daily intake level of 0.04 μg/kg bodyweight can result in an adverse analytical finding (AAF) in doping controls, healthy volunteers were administered single or multiple oral doses of 0.5μg or 3μg zilpaterol to mimic ingestion of contaminated cattle meat. Urine samples were collected and analyzed using a validated high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS) method and a newly developed chiral high-performance liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry (HPLC-APCI-MS/MS) method. Urinary peak concentrations of zilpaterol were observed for all volunteers 1.5-12.5h after ingestion, and maximum levels >5ng/mL, which would constitute an AAF in doping controls, were found after the intake of 3μg of zilpaterol on five consecutive days in one out of five study participants. Noteworthy, the enantiomeric ratio of excreted zilpaterol remained constant over time. This study provides first insights into the urinary excretion of microdosed zilpaterol. Furthermore, a method was successfully developed and applied for the separation of the zilpaterol enantiomers with mass spectrometric detection.

Qualitative Determination of Polysulfide Species in a Lithium-Sulfur Battery by HR-LC-APCI-MSn with One-Step Derivatization.

Lithium-Sulfur (Li-S) batteries are one of the most promising next-generation batteries due to their ultrahigh energy density up to 500 W h kg-1. However, despite the steady progress during the last several decades, there have been significant challenges for practical applications and commercialization. One of the major issues is controlling the lithium polysulfide (LiPS) shuttling process, which causes premature cell failure. To better understand the mechanism of the LiPS shuttling chemistry, a qualitative and quantitative analysis on polysulfide species in Li-S cell has profound significance for realizing highly efficient sulfur electrochemistry. Here we report a qualitative determination of the derivatized polysulfides in the electrolyte of a custom-made Li-S pouch cell with a high-resolution liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry method for the first time. The ionization efficiency of the methylated polysulfides was affected by the tune parameters such as the corona discharge current, the vaporizer temperature, and the source capillary temperature. It was found that the source capillary temperature was the dominant parameter to increase the peak intensity of CH3S7- ion, which was the smallest peak in the spectrum. An unusual and unique ionization pattern for methylated polysulfides detected in atmospheric pressure chemical ionization negative mode was elucidated by using first-principles calculations.

Development, validation, and estimation of measurement uncertainty for the quantitative determination of nitrosamines in Sartan drugs using liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry

In the current study, a feasible and sensitive liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry (LC-APCI-MS/MS) method was developed and validated for the determination of eight nitrosamines in Sartan active pharmaceutical ingredients (APIs). The proposed LC-APCI-MS/MS method was validated as per regulatory guidelines. The limit of detection and limit of quantification was found to be in the range of 0.32–1.58 ng/mL, 1.09–4.74 ng/mL, and the recovery was in the range of 86.4–109.6% for Sartan APIs, and 82.1–114.1% for Sartan final products, respectively. The calibration curves of all nitrosamines in the concentration range of 2–100 ng/mL were carried out, and the coefficient of determination (R2) was found to be in the range of 0.9990–0.9999. The robustness of the present method was within the acceptance (<15% RSD) for system suitability. The measurement uncertainty was determined for this method, and the expanded uncertainty was found to be in the range of 16.3–25.9% for Sartan APIs and 8.41–15.6% for Sartan drug products. The precision and recovery were found to be major sources which can induce the uncertainty during the measurement of nitrosamines in Sartan API and Drug products with the developed and validated method.

Novel QuEChERS-ultra-performance liquid chromatography–atmospheric-pressure chemical ionization tandem mass spectrometry method for the simultaneous determination of vitamin D and vitamin K in vitamin-fortified nanoemulsions

Nanoemulsion is a new vehicle for food fortification. In this study, a simple and reliable method for the simultaneous analysis of vitamins D2, D3, K1, and K2 in vitamin-fortified nanoemulsions was developed using QuEChERS (quick, easy, cheap, effective, rugged, and safe) extraction and ultra-high performance liquid chromatography–atmospheric-pressure chemical ionization tandem mass spectrometry techniques. Response surface methodology was employed to optimize the extraction parameters. The method was validated for the vitamins in terms of LOD (0.03–0.25 μg/L), LOQ (0.10–0.77 μg/L), intra-day (≤4.50%), inter-day precisions (≤6.43%), and accuracy (98.5%–108.0%). The recoveries of the vitamin-fortified nanoemulsion and yogurt were in the ranges of 104.0%–109.2% and 73.3%–85.2%, respectively. The solvent consumption and analysis time were reduced by 5.6 and 3.3 folds, respectively, rendering it superior to the traditional extraction methods established by the Association of Official Analytical Chemists and the Ministry of Food and Drug Safety.

Seminal Levels of Omentin-1/ITLN1 in Inflammatory Conditions Related to Male Infertility and Localization in Spermatozoa and Tissues of Male Reproductive System.

PurposeOmentin-1/intelectin (ITLN)1 is an adipocytokine with both anti-inflammatory and anti-oxidative stress properties, and little is known about its role in male reproduction. This study was aimed at exploring the relationships among omentin-1/ITLN1, semen parameters and F2-isoprostanes (F2-IsoPs), a maker of oxidative stress, in groups of patients affected by different pathologies. In addition, omentin-1/ITLN1 immunolocalization was assessed in ejaculated spermatozoa and in tissues of male reproductive system.Patients and MethodsSemen samples of infertile patients with varicocele (n = 27), genitourinary infections (n = 17), idiopathic infertility (n = 15) and fertile men (n = 21) were analyzed following WHO guidelines, and seminal plasma were used to determine omentin-1/ITLN1 by ELISA and F2-IsoP levels by gas chromatography/negative-ion chemical ionization tandem mass spectrometry. Omentin-1/ITLN1 was localized in human sperm and in the tissue of male reproductive system.ResultsConsidering all participants, F2-IsoP and omentin-1/ITLN1 levels were positively correlated (p = 0.000), and both these indices were negatively correlated with sperm parameters. Infertile patients showed lower sperm parameters than fertile ones; varicocele and infection groups had significantly increased levels of F2-IsoPs (both p = 0.000) and omentin-1/ITLN1 (p = 0.000 and p = 0.001, respectively). Omentin-1/ITLN1 signal was located as a spot in the connecting piece (in 43.5% of cases midpiece was also labeled) of sperm from fertile men and in cytoplasmic residue and in the entire tail in sperm of patients with varicocele and genitourinary infections. A focal omentin-1/ITLN1 immunolabelling was evident in the basal area of epididymal tubule, and a diffuse signal was present in the seminal vesicle epithelium.ConclusionSemen omentin-1/ITLN1 originates from seminal vesicles, its levels increase in inflammatory conditions and are negatively correlated with sperm parameters. For this reason, a sort of protective role of omentin-1/ITLN1 can be postulated, as this adipokine shows anti-inflammatory properties also in many other biological systems.

A Study to Validate the Relevance of Semen F2-Isoprostanes on Human Male Infertility.

F2-isoprostanes (F2-IsoPs), byproducts of arachidonic acid oxidation, are one of the most reliable indices for assessing lipid peroxidation in vivo. This study aimed at evaluating the seminal F2-IsoP level in 147 patients with different reproductive conditions (varicocele, urogenital infection, idiopathic infertility) and 45 fertile controls to establish a cut-off value discriminating physiological and pathological ranges. Semen analyses were performed following WHO guidelines; F2-IsoP levels were measured by gas chromatography/negative-ion chemical ionization tandem mass spectrometry. Considering the whole group of patients, F2-IsoPs correlated negatively with normal morphology (r = −0.283, p < 0.01), viability (r = −0.245, p < 0.01), total progressive motility (r = −0.309, p < 0.01) and rapid motility (r = −0.535, p < 0.01). The area under the ROC curve for F2-IsoP levels was 0.839, indicating a good performance of the test; the Youden index showed a cut-off value of 29.96 ng/mL. Fertile men (except one) were distributed in the group of patients with F2-IsoP level < 29.96 ng/mL. Varicocele and urogenital infection groups showed the highest levels of F2-IsoPs in semen. For the first time, a cut-off for F2-IsoPs is identified in human semen. It allows discriminating different male infertility conditions by the semen F2-IsoP amounts, as an additional parameter for clinical evaluation.

Development of a Fully Automated Analytical Platform Based on Static Headspace-Gas Chromatography-Positive Chemical Ionization-Tandem Mass Spectrometry for the Analysis of Five N-Nitrosamines in Dried Aquatic Products of Animal Origin

Development of a Fully Automated Analytical Platform Based on Static Headspace-Gas Chromatography-Positive Chemical Ionization-Tandem Mass Spectrometry for the Analysis of Five N-Nitrosamines in Dried Aquatic Products of Animal Origin

Unusual polymethylene-interrupted, Δ5 monounsaturated and omega-3 fatty acids in sea urchin (Arbacia punctulata) from the Gulf of Mexico identified by solvent mediated covalent adduct chemical ionization mass spectrometry

Sea urchin (class Echinoidea) gonads are a prized delicacy in Japan and many other world cultures. The complexity of its fatty acid (FA) profile, particularly minor FA, presents a formidable analytical challenge. We applied solvent mediated (SM) covalent adduct chemical ionization (CACI) tandem mass spectrometry to comprehensive de novo structural and quantitative characterization of the FA profile of Gulf of Mexico Atlantic sea urchin (Arbacia punctulata). >100 FA were detected including many with unusual double bond structure. Gulf sea urchin gonad lipids are rich in Δ5 monounsaturated FA 20:1(5Z) at 2.7% and the polymethylene-interrupted (PMI) diene 20:2(5Z,11Z) at 4.9%, as well as common omega-3 eicosapentaenoic acid (EPA; 5Z, 8Z, 11Z, 14Z, 17Z) at 9.8%±3.1% and arachidonic acid (AA; 5Z, 8Z, 11Z, 14Z) at 6.1%±2.1%. We propose plausible desaturation/elongation-based biochemical pathways for the endogenous production of unusual unsaturates. Unusual unsaturates may modify mammalian signaling and present novel bioactivities.