The following TLC plate (coated with silica gel) was obtained by separating five different samples using hexane as the mobile phase. D C 8,1 cm 6,5 cm B 4,4 cm A 2,3 cm Solvent front = 10 cm 9,3 cm J 7,8 cm 5,2 cm I H G 3,4 cm 2,3 cm F
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- 13. Electrospray time-of-flight mass spectrometry was used to analyze the eluate from a high-performance liquid chromatography separation. The mass spectrum of one chromatographic peak, containing a protein of unknown molecular mass, displays MH peaks at m/z = 5 654.208, 5 277.326, 4 947.590, 4 656.613, 4 397.990, and 4 166.576. Find the average molecular mass (M) of the neutral protein and its standard deviation.A student developed a chromatogram and found that the eluent from travelled 94 mm and the Zn^2+ cation travelled 24 mm. In the development of a chromatogram of a mixture of cations, the eluent front travelled 87 mm. if Zn^2+ is a cation of the unknown mixture, where will its band appear in the chromatogram?A chromatogram of a mixture of species A, B, and C provided the following data: a) Define "Retention time". b) Calculate the resolution between species A and B. c) Calculate the retention time of species B necessary to separate it from species A with a resolution of 1.5.
- The result of a TLC experiment showed that Compound X has an Rf values of 0.25 on a silica plate using 10% ethyl acetate in hexane as the mobile phase. Indicate if the Rfvalues of Compound X would be higher or lower in the following solvents: 50 % Ethyl Acetate in hexane1) Study the chromatograph (below) of a mixture of Compounds A and B, run on the GCs in the teaching labs at CU Boulder. Compound A has the shorter retention time. STAAT 61 1.11 227 RT TYPE AREA XXXX XXXX XXXX AREAS 0.009 55874 44.117 ARIHT 0.61 XX XX XX 1.11 2.27 XX XX XX TOTAL AREA=XX MUL FACTOR=XX 1. What is the retention time of compound A? Compound B? 2. Which compound is present in a larger amount? 3. Which compound has the lower boiling point? 4. What would happen to the retention times of compounds A and B if the column temperature were raised? 5. You suspect that compound B is octane. What can you do to provide supporting evidence for this hypothesis?1. For determining the %w/w of salicylamide and caffeine in an unknown sample using acetaminophen as an internal standard using a HPLC a) Is it possible to use the λmax (273 nm) of caffeine in the experiment instead of using 254 nm? Explain your reasoning.b) What are some advantages of using the Internal Standard method for quantitation over the External Standard method?c) How is the signal intensity seen in the chromatogram affected by the wavelength used for analysis?
- Two components in an HPLC separation have retention times that differ by 22 s. The first peak elutes in 10.5 min and the peak widths are approximately equal. Calculate the minimum number of theoretical plates needed to achieve an acceptable resolutionYou and another student were each given an unknown compound. Both samples contained colorless material. You each used the same brand of commercially prepared TLC plate and developed the plates using the same solvent. Each of you obtained a single spot of Rf = 0.75. Were the two samples necessarily the same substance? How could you prove unambiguously that they were ily identical using TLC?A 5.00-mL sample of blood was treated with trichloroacetic acid to precipitate proteins. After centrifugation, the resulting solution was brought to pH 3 and extracted with two 5-mL portions of methyl isobutyl ketone containing the lead-complexing agent APCD. The extract was aspirated directly into an air/acetylene flame and yielded an absorbance of 0.527 at 283.3 nm. Five-milliliter aliquots of standard solutions containing 0.400 and 0.600 ppm of lead were treated in the same way and yielded absorbances of 0.396 and 0.599. Find the concentration of lead in the sample in ppm assuming that Beer’s law is followed.
- How can l exploit the mobile phase composition to optimize my HPLC separation?Based on the chromatograms, Symmetry C18 (top) is less selective than Symmetry Shield RP18 (bottom) for the analytes given, true or false?A 5.00-mL sample of blood was treated with trichloroacetic acid to precipitate proteins. After centrifugation, the resulting solution was brought to pH 3 and extracted with two 5-mL portions of methyl isobutyl ketone containing the lead-complexing agent APCD. The extract was aspirated directly into an air/acetylene flame and yielded an absorbance of 0.569 at 283.3 nm. Five-milliliter aliquots of standard solutions containing 0.400 and 0.600 ppm of lead were treated in the same way and yielded absorbances of 0.396 and 0.599. Please calculate the concentration of lead in the sample in ppm.