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What are the general steps in developing as isocratic separation for reversed-phase chromatography with one organic solvent and temperature as variable?

Short Answer

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General steps in developing as isocratic separation.

To develop an isocratic separation for reversed-phase chromatography with one organic solvent .

Step by step solution

01

definition of general steps

To develop an isocratic separation for reversed-phase chromatography with one organic solvent.

And temperature as variables we need to change %B and temperature (T) using this steps

02

following this steps

in run Ahigh %B and low T

in rub Bhigh %B and high T

in rub C low %B and high T

in rub D low %B and low T

Using the chromatograms, determine the conditions between runs A, B, C and D.

Hence,

To develop an isocratic separation for reversed-phase chromatography with one organic solvent and temperature as variables we need to change %B and temperature (T) using this steps

in run Ahigh %B and low T

in rub Bhigh %B and high T

in rub Clow %B and high T

in rub Dlow %B and low T

Using the chromatograms, determine the conditions between runs A, B, C and D.

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Most popular questions from this chapter

A reversed-phase separation of a reaction mixture calls for isocratic elution with 48%methanol 52%water. If you want to change the procedure to use acetonitrile/water, what is a good starting percentage of acetonitrile to try?

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A known mixture of compounds A and B gave the following HPLC results:

A solution was prepared by mixing 12.49mgof Bplus 10.00mLof unknown containing just and diluting to 25.00mL. Peak areas of 5.97and 6.38were observed for AandB, respectively. Find the concentration of A(mg/mL)in the unknown.

The 鈥渞ule of three鈥 states that the retention factor for a given solute decreases approximately threefold when the organic phase increases by10%.In figure 25-12,tm=2.7min. Find k for peak 5at 40%B .predict the retention time for peak at B and compare the observed and predicted times.

  1. Use equation 25-1to estimate the length of a column required to achieve1.0104plates if the stationary phase particles size is10.5,5.0,3.0,or1.5渭尘

  2. If the retention time was 20mins on the 10.0渭尘 particle size column, what is the retention time on the 5.0,3.0,or1.5渭尘columns from part (a)? Assume that flow rate is constant for all columns.

  3. Use equation25-2to estimate the pressure of the column in (a) given that the pressure of the10.0渭尘column was4.4Mpa

  4. If the flow rate was2.0mL/min , what is the baseline width for the peaks on 10.5,5.0,3.0,or1.5渭尘columns form part (a)?

  5. Which of these column configurations would require a UHPLC instrument?

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