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Asparagus Sp.: Phytochemicals and Marketed Herbal Formulations
Published in Amit Baran Sharangi, K. V. Peter, Medicinal Plants, 2023
Vikas Bajpai, Pratibha Singh, Preeti Chandra, Brijesh Kumar
The dry plant parts of selected Asparagus species were powdered to a homogeneous size by a pulverizer and sieved through a 40-mesh sieve, respectively. The dried powder of each part (10 g) was weighed precisely and sonicated with 200 mL of 100% methanol for 30 min at room temperature using an ultrasonic water bath (53 kHz) and left for 24 hours at room temperature. Three replicates of the extraction process were carried out on each individual sample. The solution was filtered through Whatman filter paper and evaporated to dryness under reduced pressure using a rotatory evaporator (Buchi Rotavapor-R2, Flawil, Switzerland) at 40°C. Dried residue (1 mg) was weighed accurately and dissolved in 1 mL of methanol using an ultrasonicator (Bandelin SONOREX, Berlin). The solutions were filtered through 0.22 µm syringe filter (Millex-GV, PVDF, Merck Millipore, and Darmstadt, Germany). The filtrates were diluted with methanol to final working concentration and injected into the UPLC-MS/MS system for analysis.
Rapid Isolation of Lysosomes from Cultured Cells Using a Twin Strep Tag
Published in Bruno Gasnier, Michael X. Zhu, Ion and Molecule Transport in Lysosomes, 2020
Jian Xiong, Jingquan He, Michael X. Zhu, Guangwei Du
Centrifuge the pooled supernatant at 1000 g for 10 minutes at 4°C to remove cell debris, or pass them through a 0.45-µm sterile low protein binding SFCA syringe filter. Collect and transfer the supernatant and discard cell pellet. Aliquot the supernatant in new microcentrifuge tubes at 1 ml per tube and store them in a −80°C freezer.
Qualitative and Quantitative Determination of Bioactive Phytochemicals in Selected Cassia Species Using HPLC-ESI-QTOF-MS and UPLC-ESI-QqQLIT-MS/MS
Published in Brijesh Kumar, Vikas Bajpai, Vikaskumar Gond, Subhashis Pal, Naibedya Chattopadhyay, Phytochemistry of Plants of Genus Cassia, 2021
Brijesh Kumar, Vikas Bajpai, Vikaskumar Gond, Subhashis Pal, Naibedya Chattopadhyay
The dry plant parts of selected Cassia species were powdered to a homogeneous size by a pulverizer and sieved through a 40-mesh sieve, respectively. The dried powder of each part (5 g) was weighed precisely and sonicated with 200 mL of 100% methanol for 30 min at room temperature using an ultrasonic water bath (53 KHz) and left for 24 hours at room temperature. Three repeats of the extraction process were carried out on each individual sample. The solution was filtered through Whatman filter paper and evaporated to dryness under reduced pressure using rotatory evaporator (Buchi Rotavapor-R2, Flawil, Switzerland) at 40°C. Dried residues (5 mg) were used for in-vitro anti-proliferative activity screening and (1 mg) were weighed accurately and dissolved in 1 mL of methanol using ultrasonicator (Bandelin SONOREX, Berlin) for UPLC-MS/MS analysis. The solutions were filtered through 0.22 µm syringe filter (Millex-GV, PVDF, Merck Millipore and Darmstadt, Germany). The filtrates were diluted with methanol to final working concentration. 1 µL of 1mg/ml sample was injected in HPLC-MS system for qualitative analysis. Internal standard (IS) (50 µL) was spiked in final working solution of quantitative experiments, vortexed for 30s and 5 µL aliquot was injected into the UPLC-MS/MS system for quantitative analysis.
A thermodynamic study of F108 and F127 block copolymer interactions with liposomes at physiological temperature
Published in Journal of Liposome Research, 2022
Obed Andres Solis-Gonzalez, Juan Ramon Avendaño-Gómez, Aarón Rojas-Aguilar
The poloxamer molecular weight, polydispersity index (PDI), and hydrodynamic diameter (Dh) was determined by gel permeation chromatography/size exclusion chromatography (GPC/SEC) using an OMNISEC system (Malvern Panalytical, Malvern, UK). This system contains right-angle and low-angle light scattering (RALS/LALS) detectors which allow the absolute molecular weight to be determined at each time increment. From this information, the molecular weight distribution of the sample can be calculated, as well as the hydrodynamic diameter of the poloxamers. The samples were prepared in water at initial concentrations of 5.78 and 5.36 mg/mL for the F108 and F127 samples, respectively. After gentle stirring under ambient conditions for about 4 h, the samples were passed through a 0.2-µm pore-size nylon membrane syringe filter. The poloxamer aqueous solutions were analyzed using the OMNISEC GPC/SEC system with the following parameters: mobile phase, 0.05 M sodium sulphate; flow rate, 1.0 ml/min; injection volume, 100 µL; and column detector temperature, 25 °C.
A novel aqueous dimethyl trisulfide formulation is effective at low doses against cyanide toxicity in non-anesthetized mice and rats
Published in Clinical Toxicology, 2022
D. S. Lippner, D. M. Hildenberger, M. O. Rhoomes, J. N. Winborn, H. Dixon, J. McDonough, G. A. Rockwood
An isotonic aqueous formulation was prepared by the addition of 75 g Kolliphor-ELP (22 weight [wt] %) in 227 g deionized water (67 wt%), followed by the addition of 33.7 g DMTS (10 wt %) and approximately 0.7 g NaCl (0.2 wt%). The formulation was filtered through a 0.2 µm PTFE syringe filter. The concentration of DMTS in the formulation was analyzed by high-performance liquid chromatography (HPLC), and the osmolality of the formulation was analyzed on an osmometer (Model 3250, Advanced Instruments Inc.) based on freezing-point depression. The aqueous formulation was diluted in ethanol prior to HPLC analysis. The concentration of DMTS in the solution was approximately 100 mg/mL. The osmolality of the aqueous formulation was on average 300 mOsm/kg, the pH was approximately 4–6, and the solution was readily injectable through a 27–28 gauge syringe needle.
MicroRNA-mediated calcineurin signaling activation induces CCL2, CCL3, CCL5, IL8, and chemotactic activities in 4,4′-methylene diphenyl diisocyanate exposed macrophages
Published in Xenobiotica, 2021
Chen-Chung Lin, Brandon F. Law, Justin M. Hettick
MDI-GSH conjugates were prepared as previously described (Lin et al. 2020). Briefly, a 10 mM GSH solution was prepared in sodium phosphate buffer (200 mM; pH = 7.4). Fifty microlitres of freshly prepared stock solution of 10% MDI (w/v) in dry acetone were added to 25 ml of GSH solution dropwise with stirring, to a final MDI concentration of 800 µM, after which the tube was subjected to end-over-end mixing for 1 h at 25 °C. Samples were centrifuged at 10 000 × g and filtered with a 0.2 μm syringe filter. MDI-GSH conjugates were prepared immediately before use and used to treat 1 × 106 enhanced differentiated THP-1 macrophages in a 6 well plate. After 24 h, the cell culture supernatant (conditioned media) was collected, centrifuged, and stored at −20 °C until ready to use. Cells were washed 2 times with warm PBS and cell lysates were prepared by adding 600 µl of lysis/binding solution from the mirVana™ miR Isolation Kit (ThermoFisher Scientific) and stored at −80 °C until ready to use.