Question

Which is the best HPLC mobile phase solvent to use if the analytes absorb in the UV region of 210 nm? Briefly describe two other mobile phase characteristics that should be considered in HPLC analysis. List two conditions that may cause the HPLC pump pressure to become excessively high and how to resolve it without damaging the pump.

Answer 1

•Best HPLC mobile phase solvent to use if the analytes absorb in the UV region of 210 nm

Ans : UV SPECTROMETRY.

•Two other mobile phase characteristics that should be considered in HPLC analysis.

Ans: fast protein liquid chromatography( it is similar to HPLC and yeild high resolution.since FPLC needs only a relatively low backpressubac to drive the high flow rates at which separation can be performed.)

supercritical fluid chromatography( it is method which uses supercritical fluid as a mobile phase which behaves like a compressible fluid of very low viscosity where mass can be transferred very quick it also allows the use of high linear velocities.)

• two conditions that may cause the HPLC pump pressure to become excessively high and how to resolve it without damaging the pump?

Ans: 1st condition

continuous use of HPLC columns under high pressure can cause deterioration and overload of the equipment.

2nd condition

High pressure can be cause by clogged tubing i.e. salt deposit in tubing or line filter of pump is clogged or Manual injector is clogged.

To resolve it we should have to do thing like

•DISCONNECT the column and any other equipment before plumbing water through the tubing.or TAKE apart the line filter and soak it in the solvent then clean in an ultrasonic cleaner.or INJECT with 20 ml of contaminated dissolving solvent respectively in order.