Novel Fast Chromatography-Tandem Mass Spectrometric Quantitative Approach for the Determination of Plant-Extracted Phytosterols and Tocopherols

Phytosterols and tocopherols are commonly used in food and pharmaceutical industries for their health benefits. Current analysis methods rely on conventional liquid chromatography, using an analytical column, which can be tedious and time consuming. However, simple, and fast analytical methods can facilitate their qualitative and quantitative analysis. In this study, a fast chromatography-tandem mass spectrometric (FC-MS/MS) method was developed and validated for the quantitative analysis of phytosterols and tocopherols. Omitting chromatography by employing flow injection analysis—mass spectrometry (FIA-MS) failed in the quantification of target analytes due to analyte-to-analyte interferences from phytosterols. These interferences arise from their ambiguous MS fingerprints that would lead to false identification and inaccurate quantification. Therefore, a C18 guard column with a 1.9 µm particle size was employed for FC-MS/MS under isocratic elution using acetonitrile/methanol (99:1 v/v) at a flow rate of 600 µL/min. Analyte-to-analyte interferences were identified and eliminated. The false peaks could then be easily identified due to chromatographic separation. In addition, two internal standards were evaluated, namely cholestanol and deuterated cholesterol. Both internal standards contributed to the observed analyte-to-analyte interferences; however, adequate shift in the retention time for deuterated cholesterol eliminated its interferences and allowed for an accurate quantification. The method is fast (1.3 min) compared to published methods and can distinguish false peaks observed in FIA-MS. Seven analytes were quantified simultaneously, namely brassicasterol, campesterol, stigmasterol, β-sitosterol, α-tocopherol, δ-tocopherol, and γ-tocopherol. The method was successfully applied in the quantitative analysis of phytosterols and tocopherols present in the unsaponifiable matter of canola oil deodorizer distillate (CODD). β-sitosterol and γ-tocopherol were the most abundant phytosterols and tocopherols, respectively.     

The development and assessment of high‐throughput mass spectrometry‐based methods for the quantification of a nanoparticle drug delivery agent in cellular lysate

The safe use of lipid‐based drug delivery agents requires fast and sensitive qualitative and quantitative assessment of their cellular interactions. Many mass spectrometry (MS) based analytical platforms can achieve such task with varying capabilities. Therefore, four novel high‐throughput MS‐based quantitative methods were evaluated for the analysis of a small organic gene delivery agent: N,N‐bis(dimethylhexadecyl)‐1,3‐propane‐diammonium dibromide (G16‐3). Analysis utilized MS instruments that detect analytes using low‐resolution tandem MS (MS/MS) analysis (i.e. QTRAP or linear ion trap in this work) or high‐resolution MS analysis (i.e. time of flight (ToF) or Orbitrap). Our results indicate that the validated fast chromatography (FC)‐QTRAP‐MS/MS, FC‐ LTQ‐Orbitrap‐MS, desorption electrospray ionization‐collision‐induced dissociation (CID)‐MS/MS and matrix assisted laser desorption ionization‐ToF/ToF‐MS MS methods were superior in the area of method development and sample analysis time to a previously developed liquid chromatography (LC)‐CID‐MS/MS. To our knowledge, this is the first evaluation of the abilities of five MS‐based quantitative methods that target a single pharmaceutical analyte. Our findings indicate that, in comparison to conventional LC‐CID‐MS/MS, the new MS‐based methods resulted in a (1) substantial reduction in the analysis time, (2) reduction in the time required for method development and (3) production of either superior or comparable quantitative data. The four new high‐throughput MS methods, therefore, were faster, more efficient and less expensive than a conventional LC‐CID‐MS/MS for the quantification of the G16‐3 analyte within tissue culture. When applied to cellular lysate, no significant change in the concentration of G16‐3 gemini surfactant within PAM212 cells was observed between 5 and 53 h, suggesting the absence of any metabolism/excretion from PAM212 cells. Copyright © 2014 John Wiley & Sons, Ltd.     

Preparation of multiwall carbon nanotubes (MWCNTs) stabilised by highly branched hydrocarbon surfactants and dispersed in natural rubber latex nanocomposites

The performance of single-, double- and triple-chain anionic sulphosuccinate surfactants for dispersing multiwall carbon nanotubes (MWNCTs) in natural rubber latex (NR-latex) was studied using a range of techniques, including field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), thermogravimetric analysis (TGA) and Raman spectroscopy. The conductivities of the nanocomposites were also investigated using four-point probe measurements. Here, MWCNTs were efficiently dispersed in NR-latex with the aid of hyperbranched tri-chain sulphosuccinate anionic surfactants, specifically sodium 1,4-bis(neopentyloxy)-3-(neopentyloxycarbonyl)-1,4-dioxobutane-2-sulphonate (TC14). This paper highlights that TC14 performs much better than that of the commercially available surfactant sodium dodecyl sulphate (SDS), demonstrating how careful consideration of surfactant architecture leads to improved dispersibility of MWCNTs in NR-latex. The results should be of significant interest for improving nanowiring applications suitable for aerospace-based technology.     

An overview of synthetic modification of nitrile group in polymers and applications

The physicochemical properties of polymers are mainly dependent on the nature of polymer backbone and/or pendant groups linked to the main chain. Therefore, synthetic modification of these functional groups via post functionalization is an important approach for obtaining novel polymeric systems with improved properties and targeted applications. In this context, the synthetic modifications of nitrile group in polymers into various useful functionalities have received considerable attention and several interesting applications of the resulting polymers have been identified. The majority of the studies are based on Polyacrylonitrile (PAN), and some isolated examples of nitrile functionalization in copolymers such as Poly (Styrene-co-Acrylonitrile) (SAN), Poly (Acrylonitrile-co-Butadiene-co-Styrene (ABS) and Nitrile Rubber (NBR) are available. These synthetic modifications are mainly accomplished by the reactions such as Nucleophilic addition, cycloaddition, reduction, and hydrolysis using various reagents. These studies describing the post-polymerization modifications of nitrile group in polymers reported during the last three decades are covered in this review.     

Metal Enhanced Electrochemical Cyclooxygenase‐2 (COX‐2) Sensor for Biological Applications

Pain measurement is commonly required in biomedical and other emergency situations, yet there has been no pain biosensor reported in literature. Conventional approaches for pain measurement relies on Wong‐Baker face diagrams, which are grossly inadequate for situations involving children or unconscious people. We report a label‐free immunosensor for monitoring the pain biomarker cylooxygenase‐2 (COX‐2) in blood. The sensor is based on the concept of metal‐enhanced detection (MED). MED relies on the idea that the immobilization of underpotential deposition (upd) metallic films deposited either as a monolayer or electrostatically held onto a solid gold substrate could significantly amplify bimolecular recognition such as involving antigen‐antibody (Ab‐Ag) interactions. The surface bound Ab‐Ag complex insulates the electrode; causing a decrease in concentration‐dependent redox signals. A linear detection range of (3.64–3640.00)×10^?4 ng/mL was recorded with a detection limit of 0.25×10^?4 ng/mL, which was 4 orders of magnitude lower than that reported for ELISA for the same biomarker. The immunosensor exhibited selectivity of less than 6 % for potential interferents.     

Characterization of the O-4 phosphorylated and O-5 substituted Kdo reducing end group and sequencing of the core oligosaccharide of Aeromonas salmonicida ssp salmonicida lipopolysaccharide using tandem mass spectrometry

The molecular structure of the wild strain of the lipopolysaccharide core of Aeromonas salmonicida, ssp salmonicida has been sequenced using tandem mass spectrometry. The core oligosaccharide was determined to contain an O-4 phosphorylated and O-5 substituted Kdo reducing group, and its structure is proposed as the follows: [structure: see text] After the core oligosaccharide of LPS was released from the lipid A portion by conventional treatment with 1% acetic acid, we demonstrated the existence of a homogeneous mixture composed mainly of the native core oligosaccharide containing the Kdo with its O-4 phosphate group intact, and a degraded core oligosaccharide mixture, which eliminated the O-4 phosphate group with extreme facility. The precise molecular structure and glycone sequence of the homogeneous mixture of phosphorylated and dephosphorylated core oligosaccharides was determined by electrospray ionization (ESI) mass spectrometry and tandem mass spectrometric analysis. CID-MS/MS of the homogeneous mixture of permethylated core oligosaccharides afforded a series of diagnostic product ions which confirmed the established sequence of the glycones to be determined. Matrix-assisted laser desorption/ionization (MALDI) tandem mass spectrometry reconfirmed the molecular structure of the dephosphorylated homogeneous permethylated mixture of the core oligosaccharides containing the diastereomeric 4,8- and 4,7-anhydro-alpha-keto acids.     

Characterization and de novo sequencing of snow crab tropomyosin enzymatic peptides by both electrospary ionization and matrix‐assisted laser desorption ionization QqToF tandem mass spectrometry

The protein tropomyosin (TM) is a known major allergen present in shellfish causing frequent food allergies. TM is also an occupational allergen generated in the working environment of snow crab (Chionoecetes opilio) processing plants. The TM protein was purified from both claw and leg meats of snow crab and analyzed by electrospray ionization and matrix‐assisted laser desorption/ionization (MALDI) using hybrid quadruple time‐of‐flight tandem mass spectrometry (QqToF‐MS). The native polypeptide molecular weight of TM was determined to be 32 733 Da. The protein was further characterized using the ‘bottom‐up’ MS approach. A peptide mass fingerprinting was obtained by two different enzymatic digestions and de novo sequencing of the most abundant peptides performed. Any post‐translational modifications were identified by searching their calculated and predicted molecular weights in precursor ion spectra. The immunological reactivity of snow crab extract was evaluated using specific antibodies and allergenic reactivity assessed with serum of allergic patients. Subsequently, a signature peptide for TM was identified and evaluated in terms of identity and homology using the basic local alignment search tool (BLAST). The identification of a signature peptide for the allergen TM using MALDI‐QqToF‐MS will be critical for the sensitive and specific quantification of this highly allergenic protein in the work place. Copyright © 2010 John Wiley & Sons, Ltd.     

Control of the oscillator strength of the exciton in a single InGaN-GaN quantum dot

We report direct evidence for the control of the oscillator strength of the exciton state in a single quantum dot by the application of a vertical electric field. This is achieved through the study of the radiative lifetime of a single InGaN-GaN quantum dot in a p-i-n diode structure. Our results are in good quantitative agreement with theoretical predictions from an atomistic tight-binding model. Furthermore, the increase of the overlap between the electron and hole wave functions due to the applied field is shown experimentally to increase the attractive Coulomb interaction leading to a change in the sign of the biexcitonic binding energy.     

Generalized Maxwell model for the viscoelastic behavior of a soda-lime-silica glass under low frequency shear loading

The linear viscoelastic behavior of a soda-lime-silica glass under low frequency shear loading is investigated in the glass transition range. Using the time-temperature superposition technique, the master curves of the shear dynamic relaxation moduli are obtained at a reference temperature of 566°C. A method to determine the viscoelastic constants from dynamic relaxation moduli is proposed. However, some viscoelastic constants cannot be directly measured from the experimental curves and others cannot be precisely obtained due to non-linearity effects at very low frequencies. The generalized Maxwell model is investigated from the experimental dynamic moduli without fixing the viscoelastic constants. A set of parameters is shown to be in good agreement with the experimental dynamic relaxation moduli, but does not give the correct values of the viscoelastic constants of the investigated glass. The soda-lime-silica glass exhibits a non-linear viscoelastic behavior at very low stress level which is usually observed for organic glasses. This non-linear behavior is questioned.