Erlenmeyer flasks (300 mL), each containing 100 mL of the sterile cultivation medium (3% glucose, 1% peptone) (POCH, Gliwice, Poland), were aseptically inoculated with a suspension of each entomopathogenic strain and then incubated for 3 days at 24 °C on a rotary shaker. After this incubation period, 10 mg of the substrate (pregn-1,4-diene-3,20-dione (2 ) dissolved in 1 mL of dimethyl sulfoxide (DMSO) (Merck, Darmstadt, Germany)) was introduced into each flask. Sampling was conducted on the 1st, 3rd, 7th, and 10th days of the cultivation process, with the experiment being repeated in triplicate for validation. Subsequently, all reaction products were extracted using ethyl acetate, and the resulting extracts were subjected to drying with anhydrous MgSO4 (Merck, Darmstadt, Germany), followed by concentration under vacuum. The concentrated samples were then analyzed using TLC (Merck, Darmstadt, Germany), GC, and HPLC (Thermo Scientific, Waltham, MA, USA) methods, without employing sample derivatization. Quantitative analyses of the reaction mixtures were performed using GC and HPLC (Thermo Scientific, Waltham, MA, USA). This comprehensive approach ensured the accurate assessment of the biotransformation process in the tested entomopathogenic strains.
Anhydrous mgso4
Manufactured by Merck Group
Sourced in Germany, United States
Anhydrous MgSO4 is a chemical compound consisting of magnesium and sulfate. It is a fine white powder that is used as a desiccant and drying agent in various laboratory and industrial applications.
Automatically generated - may contain errors
Lab products found in correlation
Sodium chloride (nacl), by Merck Group (6 mentions)
Methanol, by Merck Group (4 mentions)
Milli q system, by Merck Group (4 mentions)
Sodium hydroxide (naoh), by Merck Group (3 mentions)
Acetonitrile, by Merck Group (3 mentions)
Gallic acid, by Merck Group (3 mentions)
Acetone, by Merck Group (2 mentions)
Acetic acid, by Merck Group (2 mentions)
Catechin, by Merck Group (2 mentions)
Ch2cl2, by Merck Group (2 mentions)
Tributyrin, by Merck Group (2 mentions)
Anhydrous sodium sulfate na2so4, by Merck Group (2 mentions)
Florisil, by Merck Group (2 mentions)
Cypermethrin, by LGC (2 mentions)
Standards of lambda cyhalothrin, by LGC (2 mentions)
Alpha hch, by LGC (2 mentions)
Beta hch, by LGC (2 mentions)
Gamma hch, by LGC (2 mentions)
Delta hch, by LGC (2 mentions)
Alpha and beta endosulfan, by LGC (2 mentions)
21 protocols using anhydrous mgso4
1
Biotransformation of Pregn-1,4-diene-3,20-dione
2
Extraction of AHLs from P. salmonis
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AHLs from P. salmonis LF-89 and Ps007 strains culture supernatants were obtained according to the method described by (Shaw et al., 1997 (link)) with some modifications. Briefly, each P. salmonis strain was grown in 600 mL of ASB at 18°C for 120 h and 75 rpm. Bacterial cells were centrifuged at 5,000 rpm for 15 min at 4°C, and the supernatant was extracted three times with a ratio of 1:1 of acidified ethyl acetate (0.5% glacial acetic acid). The organic phase obtained was dried with anhydrous MgSO4 (Merck, Darmstadt, Germany), filtered through a cellulose filter (47 mm, 0.22 µm) and concentrated by rotary evaporation at 43 ± 2°C. The dry extract was weighed and resuspended in 5 mL of acetone (GC-ECD/FID grade, Merck, Darmstadt, Germany). The extracts were stored at -20°C until use.
3
Quantification of Chlorpyrifos-methyl and Lambda-cyhalothrin Insecticides
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Insecticide (chlorpyrifos-methyl and lambda-cyhalothrin) standards were purchased from a Dr. Ehrenstorfer Lab., GmbH (Wesel, Germany) at purity of 99.69 and 98.73%, respectively; and anhydrous MgSO4, sodium acetate (NaAC), methanol (MeOH) and acetonitrile (MeCN) from Merck Co. (Darmstadt, Germany) at purities of 99.5, 99.0, 99.9 and 99.9%, respectively. PSA (primary-secondary amine 40 µm, 100 g) was sourced from Agilent (Santa Clara, CA, USA). Toxicological and physicochemical properties of insecticides are given in Table 1.
Stock solutions (400 µg/mL) were prepared adding 10 mg pesticide into 25 mL flasks and completing the final volume to 25 mL with acetonitrile. Working solutions (1.0 µg/mL) were prepared through dilution of the stock solutions. Calibration solutions were prepared over ranges of 20-4000 pg/µL and 10-2000 pg/µL. Spiking solutions equal to 0.5, 1 and 5 times of MRL were prepared. The standards and solutions were stored at 4ºC in the dark. Representative apple matrix was used for matrix-matched calibrations and quantification, as indicated in Codex Alimentarius Commission Guidelines (CAC, 2003) and SANTE Guidelines (SANTE, 2019) . Spiking with five times MRL was diluted to fit the calibration range.
Stock solutions (400 µg/mL) were prepared adding 10 mg pesticide into 25 mL flasks and completing the final volume to 25 mL with acetonitrile. Working solutions (1.0 µg/mL) were prepared through dilution of the stock solutions. Calibration solutions were prepared over ranges of 20-4000 pg/µL and 10-2000 pg/µL. Spiking solutions equal to 0.5, 1 and 5 times of MRL were prepared. The standards and solutions were stored at 4ºC in the dark. Representative apple matrix was used for matrix-matched calibrations and quantification, as indicated in Codex Alimentarius Commission Guidelines (CAC, 2003) and SANTE Guidelines (SANTE, 2019) . Spiking with five times MRL was diluted to fit the calibration range.
4
Pesticide Residue Analysis Methodology
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Pesticide standards of high purity were obtained from Dr Efrenstorfer GmbH(Augsburg, Germany), and Riedel de Haen (Seelze, Germany). This research make use of, as Pesticide standards, lambda cyhalothrin, cypermethrin, Fenarimol, permethrin, deltamethrin, fenvalerate, methomyl, imidacloprod, propoxur, bentazon, chlorpyrifos, carbaryl, 2,4 dichlorophenoxy acetic acid (2.4 D), azinphos ethyl, tetradifon, methidathion, bromofos, fenitrathion, parathion methyl, dimethoate, monocrotophos, methamidophos. Tetradecane (C14) was considered as internal standard. HPLC-grade acetonitrile, water, Acetone and acetic acid (96%) wereobtained from Merck (Darmstadt, Germany), anhydrous MgSO4(99.5%), Na citrate tribasic dehydrate (99.8%), Na citrate dibasic sesquihydrate (99.7%), NaCl (99.5%), Primary Secondary Amine (PSA) and Graphitized carbon black (GCB) were obtained from Sigma Aldrich (St Louis, MI, USA). The formic acid (88% purity) was obtained from J. T. Baker (Phillipsburg, NJ, USA). Standard solutions were obtained by appropriate dilution of the stock solutions (1000 mg.L 1 ) in Acetone (which is injected to gas chromatography) and acetonitrile (which is injected to liquid chromatography). These solutions were stored at -25•C.
5
Antioxidant and Antimicrobial Assays of Purslane
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The purslane plants were collected in July 2022 from a privte farm. The reagents, Folin reagent, gallic acid, sodium carbonate, AlCl3, methanol, acetonitrile, anhydrous MgSO4, DPPH, acetic acid, HCl, thiobarbituric acid, NaOH, phenolphthalein, and sulphuric acid, were purchased from Sigma. Muller Hinton medium (Oxiod, Basingstoke, UK). All chemicals and kits were of analytical grade.
6
Validated Standards for Polyphenol Analysis
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Standards of gallic acid (99%), 3-hydroxytyrosol (99.5%), (−)- gallocatechin (≥98%), caftaric acid (≥97%), (−)-epigallocatechin (≥95%), (+)-catechin (≥99%), (−)-epicatechin (≥95%), (−)- epigallocatechin gallate (≥95%), caffeic acid (99%), syringic acid (≥95%), coumaric acid (99%), ferulic acid (≥99%), piceatannol (99%), trans-resveratrol (≥99%), quercetin hydrate (95%), cinnamic acid (99%), quercetin 3-b-D-glucoside (≥90%), kaempferol-3-glucoside (≥99%) and malvidin-3-O- glucoside chloride (≥95%) were purchased from Sigma–Aldrich. The standard of 2-(4-hydroxyphenyl) ethanol (tyrosol) (≥99.5%) was obtained from Fluka (Buchs, Switzerland). Stock solutions of the above mentioned compounds were prepared in methanol at concentration levels of 1000 µgm l−1. Calibration standards were dissolved in the initial mobile phase of each method (LMWP or anthocyanins, respectively). HPLC-grade Acetonitrile (MeCN) and formic acid (FA) were acquired from Mallinckrodt Baker (Inc. Phillipsburg, NJ, USA). Primary-secondary amine (PSA) and octa- decylsilane (C18) were both obtained from Waters (Milford, MA, USA). Reagent grade NaCl, anhydrous Na2CO3, anhydrous MgSO4 and anhydrous CaCl2 were purchased from Sigma–Aldrich. Ultrapure water was obtained from a Milli-Q system (Millipore, Billerica, MA, USA).
7
Analytical Reagents for NMR Spectroscopy
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CH2Cl2 (HPLC purity) and anhydrous MgSO4 were from Sigma–Aldrich, as well as tributyrin (97%). The CDCl3 (isotopic purity 99.8%D) was also from Sigma–Aldrich.
8
Synthesis of Multifunctional Polymeric Nanoparticles
9
Synthesis of (1-[(4-bromomethyl)-phenyl]-1,2,2-triphenylethene conjugate
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1-[(4-bromomethyl)-phenyl]-1,2,2-triphenylethene and 4-methyl hydroxybenzoate were purchased from Rhea Biotech Company (Xian, China). Potassium carbonate, anhydrous MgSO4, sodium hydroxide, tetrabutylammonium bromide (TBAB), 1-ethyl-3-(-3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), and N-hydroxysulfosuccinimide sodium salt (sulfo-NHS) were purchased from Sigma-Aldrich. Dialysis bag (3 KDa) was purchased from Sangon Biotech Company (Shanghai, China), Ultra-4 Centrifugal Filter Devices (3 KDa) was purchased from Merck Millipore (Darmstadt, Germany). All the solvents of analytical grade were employed without more purification. ELPs40 was kindly provided by Guangya Zhang’s Group (Huaqiao university).
10
Synthesis of Organochlorosilane Precursors
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All VHPCS samples were synthesized using the same method, with a representative example for Cl3SiCH2Cl described in detail below (Fig. 1 ). A three-neck flask equipped with a dropping funnel, magnetic stirrer, and a reflux condenser was charged with dry THF (30 mL) and magnesium turnings (2.2 g, 90 mmol). Several drops of BrCH2CH2Br were added for magnesium activation and Cl3SiCH2Cl (14.7 g, 80 mmol) diluted with a small amount of THF was then added dropwise using a dropping funnel. The reaction mixture containing the MgCl2 precipitate was stirred at 60 °C for 1–3 days, and then warmed to room temperature (∼20 °C). Thereafter, ViMgCl (1.6 M in THF, 10 mL, 16 mmol) was added into filtered solution, and the mixture was stirred for 12 h. The suspension was then cooled to room temperature as about 20 °C, and LiAlH4 (1.5 g, 40 mmol) was added. The resulting slurry was then slowly warmed to 60 °C and stirred overnight at this temperature. The slurry was then cooled in an ice bath and hydrolyzed with 4 M HCl and hexane. The organic THF/hexane phase was separated from the water phase using separation funnel. Than, the solution was dried over anhydrous MgSO4 (≥99.5%, Sigma-Aldrich), and concentrated under vacuum to afford a viscous yellow oil.
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