300 microliter predictor columns often give higher recoveries than the Tricorn columns‚ with the minimum value of 66% recovery‚ even giving good recoveries using smaller salt concentrations and buffer ionic strength‚ which Tricorn cannot‚ particularly struggling during pH 5.75‚ when almost a third of the contour plots gave recoveries 40% or below. In addition‚ there were more samples taken when using the Predictor columns (9 samples per contour plot)‚ with Tricorn columns only using 5 samples per
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supernatant is to be removed and labeled accordingly in a separate vessel. Supernatant is the liquid above the sediment after a solution has been run through a centrifuge. The now centrifuged solution should be ran through a device that achieves column chromatography‚ such as an automated fraction collector. These devices sort a solution based on particle size. Other methods to extract proteins (which can be many different
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procedures of this laboratory will be used to gain a conceptual and practical understanding of separation techniques used to isolate monomers from their respective polymers. Namely‚ the techniques that will be explored are dialysis and gel filtration chromatography. In the remaining two experimental procedures‚ colorimetric assays will be used to detect the presence of certain carbohydrates. Glucose oxidase and Iodine Reactions will be performed in conjunction to demonstrate the procedure of such assays
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To purify the protein in the cell lysate from lab 1 through nickel affinity chromatography. Protein purification should result in only one type of protein ideally‚ which is the protein of interest‚ wt-DHFR and mut-DHFR in this case. A pure protein allows for further analysis on the protein to be conducted‚ such as its concentration (Bradford assay)‚ its molecular weight‚ and its biological activity. 2. Overview of experiments Buffer Preparation Add the liquid sodium phosphate‚ solid sodium chloride
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1.5.2.5 Detectors The detectors used in UPLC should be able to handle very fast scanning methods because half-height peak widths of less than one second are usually obtained with columns packed with 1.7 µm particles. The detector must be able to give high sampling rate adequate to capture enough data points across the peak for an accurate and reproducible integration of analyte peak. The dispersion (volume) of the detector flow cell must be minimal to maintain separation efficiency. Conceptually
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EXPERIMENT 4 ANALYSIS OF HYDROCARBON IN COMMON FUELS BY SOLID-PHASE MICROEXTRACTION (SPME) AND GAS CHROMATOGRAPHY-MASS SPECTROMETRY (GC-MS) Abstract In the analysis the solid-phase microextraction (SPME) and capillary gas chromatography/mass spectrometry (GC/MS) was developed for the identification of volatile compounds (hydrocarbon) in fuel. The samples was used is (kerosene‚ diesel‚ thinner and petrol) and one unknown. After the analyte was extracted by SPME in 20min‚ it directly injected
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bp). SDS-PAGE analysis of auto-induced positive clone cell lysate revealed the expression of the cfp-wzb fusion protein (experimental: 44.3 kDa). Using a hexahistidine tag‚ successful fusion protein purification was achieved via nickel affinity chromatography and confirmed by SDS-PAGE. Finally‚ para-nitrophenyl phosphate assay of tyrosine phosphatase activity allowed for determination of wzb kinetic properties such as Km (6.34 mM) and Vmax (0.0644 µmol/min/mg). Gaining an experimental competency in
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of the sample (as analyzed by 1H and 13C NMR) was poor. There are many factors that may have caused poor purity‚ of which include reaction incompletion during exposure to microwave radiation‚ poor mixing‚ and poor product isolation during column chromatography. As seen in the 1H NMR spectrum‚ impurity peaks (>10%) corresponding to methyl (triphenylphosphoranylidene) acetate‚ acetone‚ ethyl acetate‚ and hexane were identified. The phosphonium ylide peak indicates that the reaction did not go to completion
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GM‚ Kriz GS‚ Engel RG. (2011). A Small Scale Approach to Organic Laboratory Techniques. Location: Unknown. Publisher: Mary Finch (August 10‚ 2006). Retrieved November 13‚ 2012‚ from http://www.chem.ucla.edu/~bacher/General/30BL/gc/theory.html Chromatography. Introductory Theory. Retrieved November 13‚ 2012‚ from http://teaching.shu.ac.uk/hwb/chemistry/tutorials/chrom/chrom1.htm
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velmpharm24@gmail.com A precise‚ accurate and rapid RP-HPLC method has been developed and validated for the simultaneous estimation of Tamsulosin Hydrochloride and Finasteride in tablet dosage form. Chromatography was carried on RP – 18e Hiber RT (250 × 4.6) column using Water:Methanol (30:70%) v/v as mobile phase at a flow rate 0.7 ml/min and the effluent was monitored at 225 nm. The retention times of Tamsulosin Hydrochloride and Finasteride were 5.4 and 15.4 minutes respectively
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