use figure 25-17to suggest which type of liquid chromatography you could use to separate compounds in each of the following categories.

(a)Molecular mass <2000,soluble in octane

(b) Molecular mass <2000 ,soluble in methanol-water mixtures

(c) Molecular mass <2000 ,weak acid

(d)Molecular mass<2000 ,soluble highly polar

(e) Molecular mass <2000 ,ionic

(f)Molecular mass<2000,soluble in water in nonionic various

(g) Molecular mass<2000,soluble in water in water, variety of changes

(h) Molecular mass<2000,soluble in tetrahydrofuran

Short Answer

Expert verified

which type of liquid chromatography you could use to separate compounds.

a)We would use normal-phase chromatography

b)We would use bonded reverse-phase chromatography

c)We would use bonded reverse-phase chromatography.

d)We would use hydrophilic interaction chromatography

e)We would use ion-exchange or ion

f)We would use molecular-exclusion

g)We would use ion-exchange with wide pore stationary phase chromatography

h)We would use molecular-exclusion chromatography.

Step by step solution

01

Molecular mass

Figure 25-17is used to solve this.

If the analyte's molecular mass is less than 2 000 , we use the upper part of the figure. We use the lower part if the molecular mass is greater than 2 000 .Table 25-4can also be used to determine the polarity.

a)Molecular mass <2000,soluble in octane.

We would use normal-phase chromatography to separate compound that has molecular mass below 2 000 and it is soluble in octane.

b)Molecular mass <2000,soluble in methanol-water mixtures

We would use bonded reverse-phase chromatography to separate compound with molecular mass below 2 000 and it is soluble in methanol-water mixtures.

c)Molecular mass <2000,weak acid

We would use bonded reverse-phase chromatography with buffered mobile phase to separate compound with molecular mass below 2 000 and it is weak acid.

02

soluble highly polar

d)Molecular mass <2000,soluble highly polar

We would use hydrophilic interaction chromatography to separate compound with molecular mass below 2 000 and it is highly polar.

e)Molecular mass <2000,ionic

We would use ion-exchange or ion chromatography to separate compound with molecular mass below 2 000 and it is ionic.

f)Molecular mass <2000,soluble in water in nonionic various

We would use molecular-exclusion chromatography to separate compound with molecular mass greater than 2 000 and it is soluble in water, nonionic, various sized solutes.

g)Molecular mass >2000,soluble in water in water, variety of changes

We would use ion-exchange with wide pore stationary phase chromatography to separate compound with molecular mass greater than 2000 and it is soluble in water, variety of charges.

h)Molecular mass >2000,soluble in tetrahydrofuran

We would use molecular-exclusion chromatography to separate compound with molecular mass greater than 2 000 and it is soluble in tetrahydrofuran.

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Most popular questions from this chapter

Morphine and morphine 3-b-d-glucuronide were separated on two different 50 3 4.6 mm columns with 3-mm particles.61 Column A was C18-silica run at 1.4 mL/min and column B was bare silica run at 2.0 mL/min.

(a) Estimate the volume,Vm, and time,tm, at which unretained solute would emerge from each column. The observed times are 0.65 min for column A and 0.50 min for column B.

(b) Column A was eluted with 2 vol% acetonitrile in water containing 10 mM ammonium formate at pH 3. Morphine 3-β-d-glucuro-nide emerged at 1.5 min and morphine at 2.8 min. Explain the order of elution.

(c) Find the retention factor k for each solute on column A, usingtm5 0.65 min.

(d) Column B was eluted with a 5.0-min gradient beginning at 90 vol% acetonitrile in water and ending at 50 vol% acetonitrile in water. Both solvents contained 10 mM ammonium formate, pH 3. Morphine emerged at 1.3 min and morphine 3-b-d-glucuronide emerged at 2.7 min. Explain the order of elution. Why does the gradient go from high to low acetonitrile volume fraction?

(e) From Equation 25-12 in Box 25-4, estimate k* on Column B assuming S = 4 and withtm5 0.50 min.

HPLC peak should generally not have an asymmetry factor, B/A in figure 23-14,outside the range0.9-1.5

  1. Sketch the shape of a peak with an asymmetry of 1.8
  2. What might you do to correct the asymmetry?

Two peaks emerge from a reversed-phase chromatography column as sketched in the illustration.

According to Equation 23-33, resolution is given by

Resolution=N4(α-1)α(k21+k2)

where Nis plate number, αis relative retention (Equation23-20), and k2 is the retention factor for the more retained component (Equation 23-16).

(a) If you decrease the amount of organic solvent in the mobile phase, you will increase retention. Sketch the chromatogram if retention factors increase but Nand αare constant.

(b) If you change the solvent type or the stationary phase, you will change the relative retention. Sketch the chromatogram ifαincreases but Nandk1are constant.

(c) If you decrease particle size or increase column length, you can increase the plate number. Sketch the chromatogram if Nincreases by (i) decreasing particle size and (ii) increasing column length. Assume αand k2are constant.

After poisonous melamine and cyanuric acid appeared in milk in China (Box11-3) in 2008, there was a flurry of activity to develop methods to measure these substances. An analytical method for milk is to treat 1volume of milk with 9 volumes of H2O2CH3CN(20:80vol/vol)to precipitate proteins. The mixture is centrifuged for 5minto remove precipitate. The supernatant liquid is filtered through a 0.5-μmfilter and injected into a HILIC liquid chromatography column (TSK gel Amide-stationary phase) and products are measured by mass spectrometry with selected reaction monitoring (Section 22-5). Melamine is measured in positive ion mode with the transitionm/z127→85. Cyanuric acid is measured in negative ion mode with the transition m/212842.

(a) Write the formulas for the four ions and propose structures for all four ions.

(b) Even though milk is a complex substance, only one clean peak is observed for melamine and one for cyanuric acid spiked into milk. Explain why

(a) List ways in which the resolution between two closely spaced peaks might be changed.

(b) After optimization of an isocratic elution with several solvents, the resolution of two peaks is 1.2How might you improve the resolution without changing solvents or the kind of stationary phase?

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