Sample preparation for the analysis of key metabolites from cannabinoids biosynthesis in phytoplankton using GC-MS

Cannabinoids biosynthesis in phytoplankton has attracted much attention due to the rapid development of genetic tools and the optimization of genetic transformation methods in microalgae. To monitor the biosynthesis process, proper sample preparation and practical instrumental methods are needed to measure the various precursors, intermediates, cannabinoids, and their degradation products. The objective of this study was to develop a sample preparation procedure for the quantification of olivetolic acid (OA), cannabigerolic acid (CBGA), cannabidiolic acid (CBDA), tetrahydrocannabinolic acid (THCA), olivetol (OL), cannabidiol (CBD), and tetrahydrocannabinol (THC) using single-quadrupole gas chromatography-mass spectrometry (GC-MS). Isochrysis galbana was used as the model matrix. After methanol extraction, samples were purified using solid phase extraction (SPE), silylated with N-methyl-N-(trimethylsilyl)trifluoroacetamide, and analyzed using GC-MS in electron ionization mode. A strong anion-exchange SPE efficiently recovered OA, CBGA, CBDA, and THCA. A graphitized carbon black SPE was necessary to purify OL, CBD, and THC. Both columns removed amino acids, sugars, polyols, and pigments from the algae extract and prepared samples that are suitable for silylation and GC-MS analysis. The total protocol, including solvent extraction, SPE, silylation, and GC-MS analysis, was validated in accordance with the ICH guidelines. Performance characteristics of our method are superior to existing protocols with similar complexity in the literature.

Determination of 77 Multiclass Pesticides and Their Metabolitesin Capsicum and Tomato Using GC-MS/MS and LC-MS/MS

A quick, sensitive, and reproducible analytical method for the determination of 77 multiclass pesticides and their metabolites in Capsicum and tomato by gas and liquid chromatography tandem mass spectrometry was standardized and validated. The limit of detection of 0.19 to 10.91 and limit of quantification of 0.63 to 36.34 µg·kg−1 for Capsicum and 0.10 to 9.55 µg·kg−1 (LOD) and 0.35 to 33.43 µg·kg−1 (LOQ) for tomato. The method involves extraction of sample with acetonitrile, purification by dispersive solid phase extraction using primary secondary amine and graphitized carbon black. The recoveries of all pesticides were in the range of 75 to 110% with a relative standard deviation of less than 20%. Similarly, the method precision was evaluated interms of repeatability (RSDr) and reproducibility (RSDwR) by spiking of mixed pesticides standards at 100 µg·kg−1 recorded anRSD of less than 20%. The matrix effect was acceptable and no significant variation was observed in both the matrices except for few pesticides. The estimated measurement uncertainty found acceptable for all the pesticides. This method found suitable for analysis of vegetable samples drawn from market and farm gates.

Optimization of the Pre-treatment Method for the Determination of 9 Organophosphorus Pesticide Residues in Cabbage by Gas Chromatography

The purpose of this study was to optimize the pre-treatment process for the detection of 9 organophosphorus pesticide residues in cabbage by gas chromatography, and to conduct a comparative experiment between this method and the standard method NY/T 761-2008 of the Ministry of Agriculture. The results show that the optimized pre-treatment method is to use 10 mL acetonitrile as the extractant, 150 mg N-propyl ethylenediamine (PSA) and 15 mg graphitized carbon black (GCB) as the purification agent, which can reduce the use of extractant and inorganic salt in the pre-treatment process, increase the recovery and detection limit, reduce the matrix effect and cost of instrument maintenance.

Enhanced PEMFC durability with graphitized carbon black cathode catalyst supports under accelerated stress testing

The enhanced anti-corrosion properties of graphitized carbon substrates are evaluated via accelerated stress tests (ASTs). The graphitized surface properties of carbon are responsible for alleviating the coarsening and agglomeration of Pt particles.

Equilibrium, Kinetic and Thermodynamic Studies for the Adsorption of Gemfibrozil onto Graphitized Carbon Black (GCB)

Adsorption of gemfibrozil (GF) from aqueous solution by highly/lowly graphitized carbon black (GCB-H/GCB-L) was investigated by batch and fixed-bed experiments. Results showed that the adsorption of GF onto GCB greatly depended on solution pH and graphitization of carbon. The pseudo-second-order kinetic model was found to be more appropriate in describing the adsorption processes of two GCBs. The adsorption equilibriums were well fitted with the Langmuir isotherm model, and GCB-H exhibited a higher adsorption capacity of 47.68 mg g−1 at 298 K, compared with GCB-L of 4.33 mg g−1. GCB-H achieved 69.76% GF removals in simulated water after 6 h, indicating its potential practicability. The GCBs were characterized by X-ray diffraction (XRD), scanning electron microscope (SEM) and Fourier transform infrared spectroscopy (FTIR), and the results indicated that the [Formula: see text]-[Formula: see text] interaction was strongly involved in the adsorption. Thermodynamic studies further suggested that the adsorption of GF on GCB-H was exothermic, which could happen spontaneously at any temperature mainly through [Formula: see text]-[Formula: see text] interaction for physical adsorption on the surface of the plane.

Identification and Antimicrobial Activity of Medium-Sized and Short Peptides from Yellowfin Tuna (Thunnus albacares) Simulated Gastrointestinal Digestion

Due to the rapidly increasing resistance to conventional antibiotics, antimicrobial peptides are emerging as promising novel drug candidates. In this study, peptide fragments were obtained from yellowfin tuna muscle by simulated gastrointestinal digestion, and their antimicrobial activity towards Gram-positive and Gram-negative bacteria was investigated. In particular, the antimicrobial activity of both medium- and short-sized peptides was investigated by using two dedicated approaches. Medium-sized peptides were purified by solid phase extraction on C18, while short peptides were purified thanks to a graphitized carbon black sorbent. For medium-sized peptide characterization, a peptidomic strategy based on shotgun proteomics analysis was employed, and identification was achieved by matching protein sequence database by homology, as yellowfin tuna is a non-model organism, leading to the identification of 403 peptides. As for short peptide sequences, an untargeted suspect screening approach was carried out by means of an inclusion list presenting the exact mass to charge ratios (m/z) values for all di-, tri- and tetrapeptides. In total, 572 short sequences were identified thanks to a customized workflow dedicated to short peptide analysis implemented on Compound Discoverer software.

Adsorption of Sodium Hexadecyl Sulfate and Triton X from Binary Aqueous Solutions at Thermally Graphitized Carbon Black

Adsorption of anionic sodium hexadecyl sulfate (SHS) and nonionic Triton X surfactants with different ethoxylation degree: TX-45, TX-100 and TX-300 from their individual and mixed aqueous solutions at the surface of thermally graphitized carbon black (CB) was studied. It was found that at low solution concentrations addition of the nonionic surfactant increases the amount of SHS adsorbed from SHS/Triton X mixtures compared to SHS amount adsorbed from its individual solution. These findings might be explained by decreasing in electrostatic repulsion between SHS ions due to inclusion of the molecules of the nonionic surfactant in the mixed adsorption layer. At higher solution concentrations, adsorption of SHS decrease as a result of displacement of SHS ions from the mixed adsorption layer by Triton X molecules. It was established that the composition of the mixed adsorption layer at CB surface notably differ from the composition of the surfactant mixture in the bulk solution. The mixed adsorption layer is enriched with the molecules of the nonionic surfactants and this conclusion is confirmed by the results of measuring zeta potential of CB particles with the adsorbed surfactants.

Simultaneous determination of sibutramine and its derivatives in weight loss dietary supplements by LC-MS/MS

The liquid chromatography tandem mass spectrometry method (LC-MS/MS) was used to determine the content of sibutramine (SB), N-desmethyl sibutramine (DSB) and N-didesmethyl sibutramine (DDSB) illegaly mixed in weight loss dietary supplements. Sibutramine and its derivatives were extracted by methanol; impurities in the extract were removed by graphitized carbon black (GCB) adsorbent. The chromatographic separation of analytes took place on C18 column (100 mm x 2.1 mm, 3.5 µm) with a gradient mobile phase of acetonitrile and 2 mM ammonium acetate in 0.1% formic acid solution. Multiple reaction monitoring (MRM) in the positive mode was used to detect and quantify SB, DSB and DDSB at m/z 279.9/124.8; 266.0/124.8 and 252.1/125.0, respectively. The method was validated following the AOAC requirements for specificity, repeatability and recovery. Calibration curves lineared from 0.002 to 0.1 µg/mL for SB, DSB and DDSB. The method was successfully applied to determine the content of SB, DSB and DDSB in weight loss dietary supplements that were randomly collected from phamacies in Hanoi of three formulations of hard capsule, soft capsule and teabag. The results shown that six samples had SB and DSB with the content in the range of 0.817 - 31.7 mg/g.

Optimized residue analysis method for broflanilide and its metabolites in agricultural produce using the QuEChERS method and LC-MS/MS

Since broflanilide is a newly developed pesticide, analytical methods are required to determine the corresponding pesticide residues in diverse crops and foods. In this study, a pesticide residue analysis method was optimized for the detection and quantification of broflanilide and its two metabolites, DM-8007 and S(PFH-OH)-8007, in brown rice, soybean, apple, green pepper, mandarin, and kimchi cabbage. Residue samples were extracted from the produce using QuEChERS acetate and citrate buffering methods and were purified by dispersive solid-phase extraction (d-SPE) using six different adsorbent compositions with varying amounts of primary secondary amine (PSA), C18, and graphitized carbon black. All the sample preparation methods gave low-to-medium matrix effects, as confirmed by liquid chromatography–tandem mass spectrometry using standard solutions and matrix-matched standards. In particular, the use of the citrate buffering method, in combination with purification by d-SPE using 25 mg of PSA and a mixture of other adsorbents, consistently gave low matrix effects that in the range from −18.3 to 18.8%. Pesticide recoveries within the valid recovery range 70–120% were obtained both with and without d-SPE purification using 25 mg of PSA and other adsorbents. Thus, the developed residue analysis method is viable for the determination of broflanilide and its metabolites in various crops.

Multi-residue Analysis of Pesticides in Turmeric (Powder and Rhizome) Using Gas Chromatography Tandem Mass Spectrometry

Background and objectives Turmeric is widely used as an ingredient of food and medicinal products. There exists no validated method for multi-residue analysis of pesticides in turmeric. Objective This study was undertaken to develop a simple and robust method for the quantitative determination of multi-class pesticides in turmeric powder and rhizome by GC-MS/MS. Method Initially, the samples were soaked in water for 30 min and homogenized to a fine paste. A portion of this paste (2 g) was extracted with acetonitrile (2 mL) and partitioned with hexane (2 mL) after adding 5 mL of 20% NaCl. The cleanup step involved dispersive solid phase extraction with graphitized carbon black (GCB, 5 mg/mL). Its performance was evaluated against primary secondary amine (PSA) and C18 sorbents. The cleaned extract was evaporated to dryness and reconstituted in ethyl acetate before GC-MS/MS analysis. The method was validated for a mixture of 208 multi-class pesticides at 10 ng/g and higher levels (i.e., 20 and 50 ng/g). Results The findings, which demonstrated a satisfactory recovery and precision (RSDs <20%) for all compounds at 10 ng/g and higher spiking levels, are aligned with the analytical quality control criteria of SANTE/12682/2019 guidelines. The cleanup effect of GCB was much superior to that of PSA, C18, and their combinations. The solvent exchange step with hexane was effective in removing co-extractives and minimizing matrix effects. Conclusions This method complies with the regulatory requirements and is fit-for-purpose for pesticide residue monitoring in turmeric. Highlights The study reports a validated GC-MS/MS method for multi-residue analysis of pesticides in turmeric for the first time. The method provided a high throughput analysis of multi-class pesticides in turmeric rhizome and powder matrices with satisfactory selectivity, sensitivity, accuracy, and precision.