Isomeric Composition Research Articles

Fruit tree leaves as valuable new source of tocopherol and tocotrienol compounds.

Nowadays it is highly important to find new, cheap and widely available sources of tocopherol and tocotrienol compounds, and leaves are promising unconventional sources. The main goal of this study was to extend the currently limited knowledge concerning tocopherol and tocotrienol isomers composition determined using ultra-high performance liquid chromatography with fluorescence detection analysis for various fruit tree leaves such as apple, pear, quince, apricot, peach, plum, sour cherry and sweet cherry. The leaves were collected 2 weeks after tree blooming and after fruit collection. Tocopherol and tocotrienol isomers were identified and quantified for the first time in all fruit tree leaves. The total tocopherol content ranged from 203.34 to 260.86 μg g-1 dry weight for spring leaves and from 23.83 to 235.62 μg g-1 dry weight for autumn leaves and consisted mainly of α-tocopherol. The rest of the isomers of tocopherol and tocotrienols were also found, but in trace amounts. A significantly lower content of tocopherols and tocotrienols was detected in leaves after autumn collection of fruits compared to leaves collected after blooming. Among the analyzed leaves, time collected and species were significantly more important than their cultivars. Regarding quantification analysis, apricot > peach > > plums > apples leaves were identified as the best sources of tocopherols, and sweet and sour cherry leaves exhibited a lower content. Fruit tree leaves are a novel significant source and good material for isolation of α-tocopherol for application in cosmetics, pharmaceuticals or in the food industry - for example, production of beverages or other functional foods. © 2021 Society of Chemical Industry.

Stock analysis of additives synthesis via vapor-phase chromatography containing liquid crystals

Via vapor-phase chromatography 2-tiophentiol and the mixtures of individual xanthogenates, which are the stock of additive synthesis have been analyzed. Isomeric composition of high boiling aromatic hydrocarbons has been separated through vapor-phase chromatography. n,nꞌ-dyetoxiazoxybenzol and dygeptylphenyl ethers of phenyldicarbon acids, which are applied to the glass capillary columns have been used as a liquid stationary phase. The water vapor of moving carrier was used instead of the gas-carrier of nitrogen, which allowed to reduce the durability of the analysis and to improve the seperation efficiency. The main parameters of vapor-phase chromatography have been studied during the stock analysis as well. The retention time, the height of theoretic trays and rates of assimetry of analyzed compounds have been calculated.

Extract matrix composition does not affect in vitro leukotriene inhibitory effects of the Petasites hybridus extract Ze 339

It has been shown that a lipophilic CO2-extract prepared from the leaves of Petasites hybridus (Ze 339) inhibited leukotriene synthesis in vitro and ex vivo. The inhibition of the leukotriene synthesis was solely attributed to the sum of the petasins, namely petasin and its isomers isopetasin and neopetasin. To further investigate the influence of the extract matrix on leukotriene synthesis inhibition, we compared twelve selected batches of Ze 339 that differed significantly in the composition of the extract matrix. Quantitative analysis of the twelve extract batches revealed high contents of petasins [28.8–41.9%], fatty acids [17.1–27.2%] and crude oil and fat [17.7–44.2%]. The amount of sterols ranged between 3.0 and 4.9% and that of essential oils between 1.3 and 10.5%. Based on the quantitative analysis, 97–100% of the extract mass could be attributed to the above mentioned groups of ingredients. Despite significant differences in extract matrix composition, only the content of petasins was critical for the dose-dependent inhibition of leukotriene synthesis. However, at equal concentrations of petasins, no significant differences in 5-LOX, LTC4 synthase and LTA4 hydrolase inhibition were detected between the selected extract batches, despite differences in the composition of the petasin isomers. Our data suggest that the extract matrix of Ze 339 has no effect on leukotriene inhibitory effects of the petasins.

Deep Structural Annotation of Glycerolipids by the Charge-Tagging Paterno-Büchi Reaction and Supercritical Fluid Chromatography-Ion Mobility Mass Spectrometry.

Glycerolipids (GLs) are essential for cellular lipid homeostasis, while dysregulation in GL metabolism is often associated with the onset or progression of human-related metabolic diseases. The profile of GLs is thus frequently used as a molecular readout for disease phenotyping. Although mass spectrometry (MS) is the method of choice for GL profiling, the current MS methods are unable to differentiate two major types of structural isomers due to the fact that fatty acyls can be linked to different positions on the glycerol backbone (sn-positions) and the site(s) of unsaturation in acyl chains. Herein, by utilizing charge-tagging Paterno-Büchi (PB) derivatization of carbon-carbon double bond (C═C), supercritical fluid chromatography (SFC), and mobility aligned tandem mass spectrometry (MS/MS), a workflow has been developed for the sensitive and structurally informative analysis of GLs. SFC allows fast separation (within 25 min) of sn-isomers of diacylglycerols (DGs) and separation of triacylglycerols (TGs) of different chain lengths and degrees of unsaturation. Time-aligned parallel fragmentation enables multiple-stage MS/MS of the PB-derivatized lipids in a high-throughput fashion and allows pinpointing C═C location to a specific fatty acyl chain. This workflow reveals the presence of more than 500 molecular structures of neutral lipids from pooled human plasma. A comparison of human plasma samples between type 2 diabetes (N = 7) and control (N = 7) shows significant changes in isomer compositions (C18:1 Δ9 vs Δ11) from nine groups of TG and DG. These findings suggest that the developed workflow can be potentially applied to lipid marker discovery for disease monitoring or diagnosis.

In situ detection of fatty acid C=C positional isomers by coupling on-tissue mCPBA epoxidation with infrared matrix-assisted laser desorption electrospray ionization mass spectrometry.

Unsaturated fatty acids (UFAs) play vital roles in regulating cellular functions. In-depth structural characterization of UFAs such as localizing carbon-carbon double bonds is fundamentally important but poses considerable challenges in mass spectrometry (MS) given that the most widely accessible ion activation method, low-energy collision-induced dissociation (CID), primarily generates uninformative fragments (e.g., neutral loss of CO2 ) that are not suggestive of the double-bond positions. m-Chloroperoxybenzoic acid (mCPBA) was uniformly deposited onto the sample slides using a TM Sprayer, converting the carbon-carbon double bonds into epoxides under ambient conditions. The epoxidation product was ionized in situ by infrared matrix-assisted laser desorption electrospray ionization mass spectrometry (IR-MALDESI-MS), and subsequently cleaved via CID, generating a diagnostic ion pair associated with the double-bond position. The reaction efficiency, sensitivity and relative quantification capability of the method were validated with five UFA standards dried on glass slides, and then this strategy was demonstrated on thin tissue sections of rat liver and human bladder. The mCPBA reaction yielded conversion rates in the range of 44-60% in 10min with high specificity and sensitivity. Further tandem mass spectrometry (MS/MS) of the mono-epoxidized products generated informative fragment ions specific to the double-bond positions, and relative quantification of positional isomers in binary mixtures was performed across a wide mole fraction from 0 to 1. An innovative spiral scan pattern was utilized during data acquisition, elucidating the major isomeric compositions of multiple UFAs from a tissue section in a single run. The on-tissue mCPBA epoxidation was implemented into an ambient MS imaging workflow to offer a rapid and simple way for in situ identification and relative quantification of double-bond positional isomers without the requirement for instrument modification. The method can be readily implemented on many other MS platforms to reveal the role of double-bond positional isomers in lipid biology and to discover potential biomarkers.

Glass Transition Parameters of Polyisoprene Rubbers: Differential Scanning Calorimetry Study

Natural rubber and synthetic polyisoprene rubbers synthesized using titanium-, neodymium-, and gadolinium-containing catalytic systems are studied by DSC, NMR spectroscopy (700 MHz), and GPC. According to DSC thermograms, glass transition parameters, such as the temperature and width of transition and heat capacity jump, are determined. The dependence of the transition temperature on the configurational and isomeric composition of polyisoprenes with a high content of cis-units is examined. It is demonstrated that, when considering the dependence on isomeric composition, natural rubber cannot be used as a quantitative standard corresponding to the reference position of “all (100%) cis-polyisoprene” with the lowest transition temperature.

GC Analysis of Seven Seed Oils Containing Conjugated Fatty Acids

The fatty acid compositions, including isomer compositions, of seven seed oils containing conjugated fatty acids (CFA) were determined. Seed oils were extracted using a modified Folch extraction, converted to fatty acid methyl esters, and analyzed by gas chromatography (GC) with mass spectrometry and flame ionization detection. The MS detector was operated in positive-ion chemical ionization mode using methane reagent gas. GC was performed using two columns providing different retention characteristics: a poly(ethylene glycol) column and a more polar biscyanopropyl column. The complimentary information provided by the two columns was crucial to peak identification in several cases. The major CFA species in the samples are well known but all contained lesser amounts of additional CFA that have not been widely reported. All samples contained multiple species of conjugated linolenic acid, and two samples also contained small amounts of conjugated linoleic acid. The seed oils of Jacaranda mimosifolia and Calendula officinalis were found to contain 8c,10t,12t-18:3, the natural occurrence of which has only been recently reported in some other samples. The seed oil of Impatiens balsamina has been reported to contain four conjugated 18:4 species, and we present evidence for a fifth conjugated 18:4 isomer.

Assessment of organochlorine pesticides and health risk in tobacco farming associated with River Barandu of Pakistan.

Diffuse pesticide pollution through tobacco fields is a serious threat to both natural integrities and living beings because tobacco is known as a pesticide-intensive crop. Upsurge in the knowledge of detrimental impacts caused by organochlorine pesticides (OCPs) has made them a burning issue particularly in developing countries. Pakistan is a country famous for its agro-based economy and simultaneously is the second most significant pesticide consumer in South Asian countries. The studied area is tobacco hub of the country. Thus, the present work is aimed to investigate the contamination profile that highlights the ecological and health risk posed by OCPs in River Barandu, located in the proximity of tobacco farming region. ΣOCP levels in sediments ranged between 32.918 and 98.810 ng/g and in water between 0.340 and 0.935 μg/L. Hexachlorocyclohexanes (HCHs) and heptachlor were the most prevailing pesticides in both matrices of the river. Isomeric composition of DDTs and HCHs highlighted that the β-HCH and p,p'-DDT were dominant isomers in water, while α-HCH and p,p'-DDT in sediment compartment. Enantiomeric compositions of HCH and DDT indicate both recent and historic uses of these compounds in the area. Indirect contamination through nearby tobacco clusters has been depicted through spatial analysis. Ecological risk assessment based upon the risk quotient (RQ) method revealed that α-endosulfan, dieldrin, heptachlor, and ∑HCHs represent a very high level of ecological risks. The OCPs' lifetime carcinogenic and non-carcinogenic health risks associated with dermal exposure to river's water were considered nominal for surrounding populations. However, detailed ecological and health risk studies are recommended considering the bio-accumulating nature of these contaminants in the food chain.

Interplay of Aging and Lot-to-Lot Variability on the Physical and Chemical Properties of Excipients: A Case Study of Mono- and Diglycerides.

The present study investigates the chemical composition governing the physical properties of mono- and diglycerides (MDGs) at the microstructural level, as a function of aging and lot-to-lot variability. The physical structure of the MDG plays a vital role in ameliorating the emulsion stability and is widely explored in diverse research horizons related to the pharmaceutical, cosmetic, and food industries. In an effort to understand the mechanism of emulsion stabilization, physical properties were extensively evaluated in selective commercial lots to determine if there is a correlation between the chemical composition of MDG and physical properties. The solid state of the MDG samples with different aging profiles was characterized using X-ray scattering, differential scanning calorimetry, attenuated total reflection-Fourier transform infrared spectroscopy, and NMR relaxometry. Moreover, the kinetic aspect of solid-state transformation was also evaluated via treating MDG samples with a heat-cool cycle. The chemical composition of MDGs was quantified using a quantitative NMR (qNMR) method. Interestingly, the X-ray scattering results demonstrated a change in the MDG polymorphic form and an increase in the %β content as a function of aging. The increase in the %β content led to the formation of rigid crystal structures of MDG, as evident from the NMR relaxometry. Chemical quantification of isomeric composition revealed chemical composition change as a potentially critical factor responsible for the altered physical structures of MDG with respect to aging and lot-to-lot variability. The findings correlated the solid-state transformation with the change in the chemical composition of the MDG as a combined effect of aging and lot-to-lot variability. This work serves as a basis to better understand the interdependency of the physicochemical properties of MDG. Furthermore, the present work can also be used as guidance for setting up the specifications of MDG, as per the required polymorphic form for a multitude of applications.

Dietary Haematococcus pluvialis powder supplementation affect carotenoid content, astaxanthin isomer, antioxidant capacity and immune‐related gene expression in Pacific white shrimp, Litopenaeus vannamei

This study was conducted to evaluate the effect of Haematococcus pluvialis powder on astaxanthin content and isomer composition, antioxidant ability and immunity of Litopenaeus vannamei after short-term feedings. Experimental diets include a commercial diet (control) and diets containing H. pluvialis powder (HP-35 and HP-70, 35 and 70 mg of carotenoids kg−1). After feeding for 15 days, the content of astaxanthin in the muscle, cephalothorax and shell of shrimp increased significantly with increasing addition of H. pluvialis powder (p < 0.05). Dietary addition of H. pluvialis powder significantly increased the activity of total antioxygenic capacity (T-AOC), superoxide dismutase (SOD) and glutathione S-transferase (GST) as well as the relative mRNA levels of immune-related genes, Toll, MyD88, IMD, Crustin, PO and Lysozyme in hepatopancreas (p < 0.05). The proportion of astaxanthin isomers in different body parts was different, whereas it has no significant difference between same body parts of different groups. This indicated that the accumulation of astaxanthin among body parts may be accompanied with selection and isomerization. In conclusion, feeding with H. pluvialis powder diets containing 35 and 70 mg of carotenoids kg−1 can enhance the carotenoid and astaxanthin content, antioxidant and immune capacity of L. vannamei.

КОНФИГУРАЦИОННО-ИЗОМЕРНЫЙ СОСТАВ КАУЧУКОВ: ОПРЕДЕЛЕНИЕ МЕТОДОМ СПЕКТРОСКОПИИ ЯМР 1H И 13C

Nuclear magnetic resonance (NMR) spectroscopy is a convenient method for studying the structure and isomeric composition of diene rubbers. A simple and reproducible method is proposed for determining the content of both rubber and impurity compounds, which does not require additional sample preparation. Formulas for calculating the composition of industrial products and determining the initial substances are given, the applicability of the analysis method is confirmed. In the process of work, rubbers of various brands were analyzed, their configuration-isomeric composition was established. It was found that the integration ranges given in the literature earlier and the methods for processing 1H NMR spectra differ, the signals of the groups often overlap in the spectrum, which increases the calculation error. It is shown that the structure of the industrial products under study is established most accurately by carbon proton resonance spectroscopy, and the calculation based on the PMR spectra allows confirming the data obtained with sufficient accuracy.

Photo-Isomerization Kinetics of Azobenzene Containing Surfactant Conjugated with Polyelectrolyte.

Ionic complexation of azobenzene-containing surfactants with any type of oppositely charged soft objects allows for making them photo-responsive in terms of their size, shape and surface energy. Investigation of the photo-isomerization kinetic and isomer composition at a photo-stationary state of the photo-sensitive surfactant conjugated with charged objects is a necessary prerequisite for understanding the structural response of photo-sensitive complexes. Here, we report on photo-isomerization kinetics of a photo-sensitive surfactant in the presence of poly(acrylic acid, sodium salt). We show that the photo-isomerization of the azobenzene-containing cationic surfactant is slower in a polymer complex compared to being purely dissolved in aqueous solution. In a photo-stationary state, the ratio between the trans and cis isomers is shifted to a higher trans-isomer concentration for all irradiation wavelengths. This is explained by the formation of surfactant aggregates near the polyelectrolyte chains at concentrations much lower than the bulk critical micelle concentration and inhibition of the photo-isomerization kinetics due to steric hindrance within the densely packed aggregates.

Spatial Trends of Anionic, Zwitterionic, and Cationic PFASs at an AFFF-Impacted Site.

Soil and groundwater from an aqueous film-forming foam (AFFF)-impacted site were sampled at high resolution (n = 105 for soil, n = 58 for groundwater) and analyzed for an extensive list of anionic, zwitterionic, and cationic poly- and perfluoroalkyl substances (PFASs). Spatial trends for perfluoroalkyl acids and many precursors enabled a better understanding of PFAS composition, transport, and transformation. All PFASs without analytical standards were semi-quantified. Summed PFAS and individual PFAS concentrations were often higher at depth than near the surface in soil and groundwater. Zwitterionic and cationic compounds composed a majority of the total PFAS mass (up to 97%) in firefighter training area (FTA) soil. Composition of PFAS class, chain length, and structural isomers changed with depth and distance from the FTA, suggesting in situ transformation and differential transport. The percentage of branched perfluorooctane sulfonate increased with depth, consistent with differential isomeric transport. However, linear perfluorooctanoic acid (PFOA) was enriched, suggesting fluorotelomer precursor transformation to linear PFOA. Perfluorohexane sulfonamide, a potential transformation product of sulfonamide-based PFASs, was present at high concentrations (maximum 448 ng/g in soil, 3.4 mg/L in groundwater). Precursor compounds may create long-term sources of perfluoroalkyl acids, although many pathways remain unknown; precursor analysis is critical for PFAS fate and transport understanding.

Chiral Analysis of Amphetamine and Methamphetamine in Urine by Liquid Chromatography-Tandem Mass Spectrometry Applying Mosher Derivatization

Methamphetamine (MA) and amphetamine (AM) are central nervous system doping agents, listed in the WADA Prohibited List, which in the past years showed the highest prevalence among all stimulants. Specifically, concerning these stimulants, it is essential to distinguish between d and l-isomers of AM and MA due to their distinction in illicit or legal nonprescription use. The main separation methods used for the urinary determination of MA and AM enantiomers are based on chromatographic procedures, such as gas chromatography after chiral derivatization or liquid chromatography (LC) applying chiral columns, followed by the mass spectrometer (MS) detection. The use of the optically pure reagent R-(-)-α-methoxy-α-(trifluoromethyl) phenylacetic acid chloride (R-MTPA-Cl – Mosher’s Reagent) allows the isomeric separation by both chromatographic procedures while preserving the isomeric composition of the substances. Therefore, in the present work, an LC-MS2 approach has been developed for AM and MA detection in urine after Mosher derivatization. The method was validated and can be used for the enantioselective confirmation of AM and MA for doping control purposes and forensic analysis in general, being useful for the distinction between the medicinal and illicit use of both stimulants.

Resolving Modifications on Sphingoid Base and N-Acyl Chain of Sphingomyelin Lipids in Complex Lipid Extracts.

Sphingomyelins (SMs) are an essential class of lipids widely existing in different organisms. The sphingoid base and N-acyl chain are two building blocks which can undergo different types of modifications during lipogenesis, including desaturation, hydroxylation, and methyl branching. Current lipidomic analysis methods cannot provide detailed information on these structural motifs. Herein, we developed a tandem mass spectrometric method by harnessing radical-directed dissociation (RDD) from collision-induced dissociation (CID) of the bicarbonate anion adduct of SM ([M + HCO3]-). A major RDD channel produced a high-abundance fragment carrying the intact N-acyl chain, termed as "N-acyl fragment", allowing the assignment of the sphingoid base/N-acyl composition and relative quantitation of compositional isomers of SM at high sensitivity. RDD also produced intrachain fragments in lower abundances, which helped localization of methyl branching and hydroxylation in SM. The acetone Paternò-Büchi (PB) reaction was found to be capable of derivatizing the Δ4 carbon-carbon double bond (C═C) in sphingosine (SPH) base and producing C═C diagnostic ions upon CID, albeit at much lower efficiencies than those of the isolated C═C in alkyl chains. A liquid chromatography-mass spectrometry workflow was developed by incorporating MS2 CID of SM via [M + HCO3]- and PB-MS2 CID. The capability of profiling SM with detailed structural information was demonstrated by analyzing complex lipid extracts from porcine brain and Caenorhabditis elegans. These results provided visualization of the sphingoid base/N-acyl compositional isomers of SM lipids and revealed large structural diversity from each sample. These included identification of the sphingadiene base [d18:1(Δ4,14)], C═C location isomers in N-acyls, C-2 hydroxylation of N-acyls, and iso-methyl branched SPH base.

Absorption and Emission Spectroscopic Investigation of the Thermal Dynamics of the Archaerhodopsin 3 Based Fluorescent Voltage Sensor Archon2.

Archon2 is a fluorescent voltage sensor derived from Archaerhodopsin 3 (Arch) of Halorubrum sodomense using robotic multidimensional directed evolution approach. Here we report absorption and emission spectroscopic studies of Archon2 in Tris buffer at pH 8. Absorption cross-section spectra, fluorescence quantum distributions, fluorescence quantum yields, and fluorescence excitation spectra were determined. The thermal stability of Archon2 was studied by long-time attenuation coefficient measurements at room temperature (21 ± 1 °C) and at refrigerator temperature (3 ± 1 °C). The apparent melting temperature was determined by stepwise sample heating up and cooling down (obtained apparent melting temperature: 63 ± 3 °C). In the protein melting process protonated retinal Schiff base (PRSB) with absorption maximum at 586 nm converted to de-protonated retinal Schiff base (RSB) with absorption maximum at 380 nm. Storage of Archon2 at room temperature and refrigerator temperature caused absorption coefficient decrease because of partial protein clustering to aggregates at condensation nuclei and sedimentation. At room temperature an onset of light scattering was observed after two days because of the beginning of protein unfolding. During the period of observation (18 days at 21 °C, 22 days at 3 °C) no change of retinal isomer composition was observed indicating a high potential energy barrier of S0 ground-state isomerization.

Semi-centennial sediment records of HCHs and DDTs from the East China marginal seas: Role of lateral transport in catchment

We reconstructed the history of the inputs of hexachlorocyclohexanes (HCHs) and dichlorodiphenyltrichloroethanes (DDTs) into the marine environment to reveal the time-dependent influence of sources and associated anthropogenic activities in China, based on 210Pb-dated cores from the East China marginal seas (ECMS). The temporal dynamics of pesticide contamination expresses as deposition fluxes, inventories, and half-life estimations varied among the cores, suggesting heterogeneity in transport pathways of pollutants. The depth profiles of pesticide inputs closely followed their historical production and application timelines in China, and were also affected by human activities in catchments, with general declines in HCH and DDT inputs to the coring sites after their peak deposition. Despite the prevalence of occurrence of weathered HCH/DDT in the cores, there were clear source-dependent differences in isomeric composition and accumulation between before and after these pesticides were banned. α-HCH and p,p′-DDT were relatively more enriched in sediments from the pre-ban period when heavy technical HCHs and DDTs use occurred, as indicated by the higher α-/β-HCH and lower (DDE + DDD)/DDTs ratios, and the larger fractions of α-HCH and p,p′-DDT influxes to the coring sites in the ΣHCH and ΣDDT fluxes, respectively, while this pattern shifted to be historical residue-based in the post-ban period. The difference in the recent influxes of pesticides to core sediments and their higher post-ban inventories highlight the increasing importance of historical sources over time and continuous input of weathered residues into marine environment via lateral transport.

Direct Analysis of Native N-Linked Glycans by IR-MALDESI.

Glycan analysis by mass spectrometry has rapidly progressed due to the interest in understanding the role of glycans in disease and tumor progression. Glycans are complex molecules that pose analytical challenges due to their isomeric compositions, labile character, and ionization preferences. This study sought to demonstrate infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) as a novel approach for the direct analysis of N-linked glycans. The glycoprotein bovine fetuin was chosen for this analysis as its glycome is well-characterized and heavily composed of sialylated glycans. Native N-linked glycans produced by enzymatic cleavage (via PNGase F) of bovine fetuin were analyzed directly by IR-MALDESI in both positive and negative ionization mode. In this study, we detected 12 N-linked glycans in negative mode and 4 N-linked glycans in positive mode, a significant increase in the amount of underivatized glycans detected by other ionization sources. Importantly, all N-linked glycans detected contained at least one sialic acid residue, which are known to be labile. This work represents a critical first step for N-linked glycan analysis by IR-MALDESI with future efforts directed at mass spectrometry imaging.

Changes in Vitamin E and β-Carotene Contents in Various Edible Cassava Leaves (Manihot esculenta Crantz) of Different Ages across Multiple Seasons

Vitamin E and carotenoids belong to a group of bioactive compounds that have an important effect on human health. The present study aims to investigate for the first time the concentration of vitamin E, α-, γ-, and δ-tocopherol, and β-carotene in edible cassava leaves during different ages. The analysis was performed using colorimetry and high-performance liquid chromatograph (HPLC) methods. A significant difference was found among α-, γ-, and δ-tocopherol concentrations in leaves; γ- and α-tocopherol isomer contents were the predominant in amount, respectively. Among the leaves, AD variety harvested at 6 months after planting (MAP) was the highest in total vitamin E (222 μgα-TE/g). However, the highest γ-tocopherol content (2782 μg/100g) and the content of biologically active vitamin E (1244 μg/100g) were found in EN variety at 6MAP, whereas the highest value (42 μg/g) of β-carotene was found in AD variety at 12MAP. Total vitamin E and tocopherol isomers composition varied among varieties and seasons as did β-carotene. The effects of varieties and harvest ages on the biosynthesis regulation of these compounds were confirmed by principal component analysis (F1x F2: 74.34%). Edible cassava leaves can be considered as sources of vitamin E for natural dietary antioxidant during different ages, and the best time to harvest EN and AD varieties is at 6MAP on the basis of α-tocopherol equivalent or content of biologically active vitamin E content, and EN and AD varieties performed the overall best. Thus, the edible cassava leaves are among the leafy vegetables that could be helpful to suggest as a part of daily meal for health benefits and pharmaceutical purposes.

Chlorines Are Not Evenly Substituted in Chlorinated Paraffins: A Predicted NMR Pattern Matching Framework for Isomeric Discrimination in Complex Contaminant Mixtures.

Chlorinated paraffins (CPs) can be mixtures of nearly a half-million possible isomers. Despite the extensive use of CPs, their isomer composition and effects on the environment remain poorly understood. Here, we reveal the isomeric distributions of nine CP mixtures with single-chain lengths (C14/15) and varying degrees of chlorination. The molar distribution of CnH2n+2–mClm in each mixture was determined using high-resolution mass spectrometry (MS). Next, the mixtures were analyzed by applying both one-dimensional 1H, 13C and two-dimensional nuclear magnetic resonance (NMR) spectroscopy. Due to substantially overlapping signals in the experimental NMR spectra, direct assignment of individual isomers was not possible. As such, a new NMR spectral matching approach that used massive NMR databases predicted by a neural network algorithm to provide the top 100 most likely structural matches was developed. The top 100 isomers appear to be an adequate representation of the overall mixture. Their modeled physicochemical and toxicity parameters agree with previous experimental results. Chlorines are not evenly distributed in any of the CP mixtures and show a general preference at the third carbon. The approach described here can play a key role in understanding of complex isomeric mixtures such as CPs that cannot be resolved by MS alone.