Fisher optima

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom

The Fisher Optima is a high-performance laboratory centrifuge designed for a wide range of applications. It features a robust construction, intuitive controls, and advanced safety features to ensure reliable and efficient operation in the laboratory setting.

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7 protocols using fisher optima

Dehydration-induced Sugar Metabolism in Tripogon loliiformis

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Tripogon loliiformis shoots at the various dehydration points described above, were sampled and analysed for sugar content by Gas Chromatography-Mass Spectroscopy (GCMS). Approximately, 100 mg of dry tissue was analysed using a Waters Acquity UPLC coupled to a mass spectrometer. For quantitation purposes, stachyose standards were analysed alongside the samples. Chromatographic separation was performed using a Waters BEH Amide UPLC column (1.7 mm, 2.1 mm 3 150 mm; 30C). The mobile phase used a mixture of water (Fisher Optima) containing 0.1% formic acid (solvent A; Fisher) and acetonitrile (Fisher Optima) containing 0.1% formic acid (solvent B) in a linear gradient from 80% B to 50% B at a flow rate of 0.35 ml/min. For statistical significance, triplicate samples were measured^{19 (link)}.

Okemo P., Long H., Cheng Y., Mundree S, & Williams B. (2021). Stachyose triggers apoptotic like cell death in drought sensitive but not resilient plants. Scientific Reports, 11, 7099.

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Metabolite Analysis by GC-MS

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All reagents were Fisher Optima grade (Fisher Scientific, Hanover Park, IL). Authentic chemical standards were obtained from Sigma-Aldrich (St. Louis, MO) (citric acid, lactic acid, cis-aconitic acid, isocitric acid, malic acid, phenylacetic acid, hippuric acid, hexanedioic acid, threonine, palmitic acid, methylsuccinic acid, succinic acid, glycine, leucine, asparagine, isoleucine, serine, lysine, pyroglutamic acid, pipecolic acid, 3-hydroxybutyric acid, glyceric acid, alanine, and aspartic acid), Santa Cruz Biotechnology (Dallas, TX) (propionylglycine), or MP Biomedicals (Santa Ana, CA) (4-hydroxyproline). Internal standards were obtained from Sigma-Aldrich (4-nitrobenzoic acid) or Cambridge Isotope Laboratories (Andover, MA) (citric acid-d₄, D-sorbitol-¹³C₁, D,L-glutamic acid-d₅, and L-leucine-d₃). Chemicals for derivatization and analysis were obtained from Sigma-Aldrich (urease type III, anhydrous sodium sulfate, C4–C24 FAMEs, methyl nonanoate, and alkane standard mix [C₁₀-C₄₀]) or Thermo Scientific (Waltham, MA) (N-methyl -N- [trimethylsilyl] -trifluoroacetamide (MSTFA) with 1% [vol/vol] trimethyl-chlorosilane (TMCS)).

Pannkuk E.L., Laiakis E.C., Authier S., Wong K, & Fornace AJ J.r. (2017). Gas Chromatography/Mass Spectrometry Metabolomics of Urine and Serum from Nonhuman Primates Exposed to Ionizing Radiation: Impacts on the Tricarboxylic Acid Cycle and Protein Metabolism. Journal of proteome research, 16(5), 2091-2100.

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Microcystin Toxin Analysis in Water Samples

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In triplicate, 200 mL of water sample was filtered (CFC, Whatman, Maidstone, UK), filter papers were wrapped individually in aluminium foil and preserved at −80 °C. On analysis, filter papers were subjected to three cycles of freeze-thawing before submersion in 10 mL of 80% aqueous methanol. Samples were left in the dark at 4–6 °C for 24 h, before ~0.5 mL was aliquoted into a LCMS certified vial. Toxin analysis was carried by ultra-high-performance liquid chromatography (UHPLC) (Acquity, Waters, Manchester, UK) coupled to a tandem quadruple mass spectrometer (Xevo TQ, Waters, Manchester, UK). All instrument solvents and chemicals were of LC-MS-grade (Fisher Optima, Thermo Fisher, Manchester, UK). Reference toxins used for the detection method included the microcystin analogues MC-RR, MC-LA, MC-LY, MC-LF, MC-LW, MC-YR, MC-WR, MC-HilR, MC-HtyR, MC-LR & Asp³-MC-LR (Enzo Life Sciences, Exeter, UK) and [Dha⁷]-MC-LR and matrix reference material of blue-green algae (RM-BGA, Lot 201301) containing a range of microcystins (Institute of Biotoxin Metrology, National Research Council Canada). Analysis of microcystins was conducted following the method by Turner et al. [50 (link)].

Hartnell D.M., Chapman I.J., Taylor N.G., Esteban G.F., Turner A.D, & Franklin D.J. (2020). Cyanobacterial Abundance and Microcystin Profiles in Two Southern British Lakes: The Importance of Abiotic and Biotic Interactions. Toxins, 12(8), 503.

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Quantitative Analysis of Cyanotoxin Standards

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Mobile phases were prepared from LC-MS-grade acetonitrile (Fisher Optima, ThermoFisher, Greater London, UK) and water used for LC-MS was obtained in-house. Sample preparation reagents were HPLC grade. Toxin standards used for preparation of calibration solutions (MC-RR, MC-LA, MC-LY, MC-LF, MC-LW, MC-YR, MC-WR, MC-HilR, MC-HtyR, MC-LR, [Asp3] MC-LR and Nodularin) were all obtained from Enzo Life Sciences, Exeter, UK. A certified standard of [Dha7] MC-LR was obtained from the Institute of Biotoxin Metrology, National Research Council Canada (NRCC, Halifax, NS, Canada). Reference standards received as solid films were dissolved in 50% aqueous methanol, to form stock solutions. A mixed stock solution was subsequently prepared by combining aliquots of each stock, followed by further dilutions to create seven-level suite of working calibration standards between 0.33 ng/mL to 327 ng/mL per toxin.

Turner A.D., Dhanji-Rapkova M., O’Neill A., Coates L., Lewis A, & Lewis K. (2018). Analysis of Microcystins in Cyanobacterial Blooms from Freshwater Bodies in England. Toxins, 10(1), 39.

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Cyanotoxin Analysis with Mass Spectrometry

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Instrument solvents used for preparation of mobile phases were of LC-MS-grade (Fisher Optima, ThermoFisher, London, UK) and all chemicals were LC-MS reagent grade where possible, otherwise HPLC grade. Sample preparation reagents were all HPLC grade. Cyanotoxin reference toxin standards comprising the microcystin analogues (MC-RR, MC-LA, MC-LY, MC-LF, MC-LW, MC-YR, MC-WR, MC-HilR, MC-HtyR, MC-LR, [Asp3] MC-LR), Nodularin (Nod), ATX, Cylindrospermopsin (CYN) were all obtained from Enzo Life Sciences (Enzo, Exeter, UK), with secondary calibration check standards and Saxitoxin (STX) standard prepared using certified reference solutions from Biotoxin Metrology, National Research Council of Canada (NRCC, Halifax, Nova Scotia, Canada). Enzo reference standards received as solid powders were dissolved in suitable volumes of 50% aqueous methanol, to form stock solutions. A mixed stock solution was prepared by combining aliquots of each stock, followed by a seven-level suite of working calibration solvent standards resulting in a calibration range between 1.0 ng/mL to 500 ng/m per toxin. Phe was also obtained from Merck (Poole, UK) and used for confirmation of resolution from ATX, given the isobaric nature of the two compounds and the potential for false positive detection of ATX in matrices containing Phe [62 (link)].

Turner A.D., Turner F.R., White M., Hartnell D., Crompton C.G., Bates N., Egginton J., Branscombe L., Lewis A.M, & Maskrey B.H. (2022). Confirmation Using Triple Quadrupole and High-Resolution Mass Spectrometry of a Fatal Canine Neurotoxicosis following Exposure to Anatoxins at an Inland Reservoir. Toxins, 14(11), 804.

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Quantification of Marine Neurotoxins

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Sample preparation and SPE reagents as well as Pre-COX HPLC-FLD reagents were HPLC grade.
Water, acetonitrile and methanol (HPLC grade), glacial acetic acid, formic acid (Laboratory grade) were purchased from Sigma-Aldrich Corporation (St. Louis, MO). Solvents and additives for UHPLC-HILIC-MS/MS and HPLC-HILIC-HRMS analysis were LC-MS grade (Fisher Optima, ThermoFisher, UK).
Certified reference material (CRM) of STX, GTX1-6, NEO, dcSTX, C1&2, dcNEO and dcGTX2&3 and noncertified but well characterised reference material of C3&4 and dcGTX1&4 were all obtained from the Institute of Biotoxin Metrology, National Research Council of Canada (NRCC, Halifax, Canada). TTX CRM was obtained from Cifga (Lugo, Spain). A reference standard for deoxydecarbamoylsaxitoxin and a freeze-dried naturally contaminated tissue of the sea slug Pleurobranchaea maculata containing extremely high concentrations of TTX and its analogues was purchased from Cawthron Natural Compounds (CNC; Nelson, New Zealand). Preparation of toxin calibration standards for PST analysis was conducted for UHPLC-HILIC-MS/MS (Turner et al., 2015c) , PreCOX HPLC-FLD (Anon, 2005c) , and for TTXs following Turner et al. (2017) .

Dell'Aversano C., Tartaglione L., Polito G., Dean K., Giacobbe M., Casabianca S., Capellacci S., Penna A, & Turner A.D. (2019). First detection of tetrodotoxin and high levels of paralytic shellfish poisoning toxins in shellfish from Sicily (Italy) by three different analytical methods. Chemosphere, 215.

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Cyanotoxin Quantification Protocol

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Instrument solvents used for preparation of mobile phases were of LC-MS-grade (Fisher Optima, ThermoFisher, UK) and all chemicals were LC-MS reagent grade where possible. Sample preparation reagents were HPLC grade. Reference toxin standards (MC-RR, MC-LA, MC-LY, MC-LF, MC-LW, MC-YR, MC-WR, MC-HilR, MC-HtyR, MC-LR, [Asp3] MC-LR and Nod) were all obtained from Enzo Life Sciences, Exeter, UK. A certified standard of [Dha 7 ]-MC-LR and a pre-certified freeze-dried matrix reference material of blue-green algae (RM-BGA, Lot 201301) containing a range of MC was obtained from the Institute of Biotoxin Metrology, National Research Council Canada (NRCC). Reference standards received as solid powders were dissolved in suitable volumes of 50% aqueous methanol, to form stock solutions. A mixed stock solution was subsequently prepared by combining aliquots of each stock, followed by a seven-level suite of working calibration standards resulting in a calibration range between 0.33 ng/mL to 327 ng/m per toxin. RM-BGA (280 mg) was extracted with 28.0 mL 50% aqueous MeOH + 0.1% acetic acid, prior to centrifugation (4,500 g; 10 min) and the supernatant collected prior to analysis. The seven-point calibration standards were used for external calibration of cyanotoxins in all sample matrices, adjusting dilution factors depending on the extraction applied.

Turner A.D., Waack J., Lewis A., Edwards C, & Lawton L. (2018). Development and single-laboratory validation of a UHPLC-MS/MS method for quantitation of microcystins and nodularin in natural water, cyanobacteria, shellfish and algal supplement tablet powders. Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 1074-1075.

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