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- You must separate three different compounds with reversed phase HPLC. Compound 1 is an acid (pKa = 4.3), compound 2 is neutral compounds and compound 3 is a base (pKa = 9.6). As eluent, use 30/70 (v / v)% methanol / pH 3.0 formic acid buffer. The retention time and the top width at the base of the top for the three components are presented below. You can assume the dead time to 1.2 minutes in your calculations.Compound Retention time [minutes] Peak width [seconds] 1 10.1 282 5.0 143 6.1 17a) Calculate the efficiency (number of theoretical bases) for component 1.b) Calculate the selectivity and resolution between components 1 and 2.c) When you increase the pH of your eluent to pH 6, how will the retention be affected (increase, decreaseor unchanged) for the different associations (1, 2 and 3) ?, justify your answer.d) How retention factors are generally affected by an increase in the proportion organicallymobile phase solvents for reversed phase HPLC separations ?, justify your…Gradient elution used in HPLC. How is the gradient created for a reverse-phase HPLC experiment over the course of the run? A)Increase the pH of the mobile phase. B)Decrease the pH of the mobile phase. C)Increase the vapor pressure of the analytes by increasing the temperature. D)Increase mobile phase polarity by increasing the fraction of polar solvent in a binary mobile phase. E)Decrease mobile phase polarity by mixing with a less polar solvent to better compete with the nonpolar stationaryphase. Plz explain properly why statement is not correctGradient elution used in HPLC. How is the gradient created for a reverse-phase HPLC experiment over the course of the run? A)Increase the pH of the mobile phase. B)Decrease the pH of the mobile phase. C)Increase the vapor pressure of the analytes by increasing the temperature. D)Increase mobile phase polarity by increasing the fraction of polar solvent in a binary mobile phase. E)Decrease mobile phase polarity by mixing with a less polar solvent to better compete with the nonpolar stationaryphase.
- Metal ions chelate polar groups of analyte molecules for longer retention time. Does this affect chromatogram of RPHPLC?Which statement about TLC analyses is not true? * A- The line at the base of the TLC plate should be drawn with a pencil. B- To prevent contamination, a new capillary tube should be used to apply each sample. C- Samples should be applied multiple times in the same place to increase the concentrations of any possible compounds in the spot. D- A glass beaker lined with filter paper can be used as a development chamber in a TLC analysis. E- All these statements are correct.27. What exactly is meant by reversed phase? In addition to finding what compound would elute first in a reverse phase HPLC Separation... Can you explain what would elute first for HPLC, GC, TLC, IEC, SEC (or gel filtration) and affinity chromatography?