Ultra Performance Liquid Chromatography (UPLC)

 

Introduction:

UPLC refers to Ultra Performance Liquid Chromatography. UPLC is an advanced mode of chromatography used in analytical technique. High performance liquid chromatography (HPLC) has proven to one of the most and predominant technology used in analytical laboratories for the analysis of drugs worldwide during the past 30- plus years. But now UPLC is replacing HPLCs to complete the analysis in short time. Ultra Performance liquid chromatography (UPLC) applicable for particle less than 2μm in diameter packing materials.

UPLC improves the analysis in the below mentioned 3 ways when compared to HPLC are,

1.  Better Resolution
2. Speed
3. Sensitivity

This technology takes full advantage of chromatographic principles to run separations using columns packed with smaller particles and high flow rates.

Working Principle:

The principle is based on Van Deemter Equation, which is familiar with chromatographic users,

                                      H=A+B/v +Cv

The above equation describes the relationship between linear velocity (flow rate) and plate height (HETP, or column efficiency). And, since particle size is one of the variables, a Van Deemter curve can be used to investigate chromatographic performance. Where A, B and C are constants and v is the linear velocity, the carrier flow rate.

                        A-Eddy diffusion

                        B-Longitudinal diffusion

                        C-Equilibrium mass transfer

"The Principle of HPLC and UPLC is same but not the Performance."

The principles of UPLC are same principle as HPLC, the basic difference is in designer of the column material particle size which less than 2-μm. Which make a big deference in performance and to maximize the advantages of these columns, creating a powerful, robust and reliable solution.

Advantages of UPLC:

1. Decrease the runtime and enhanced the sensitivity.
2. Provides the selectivity, sensitivity and dynamic range of LC analysis.
3. Resolved peaks are obtained in chromatogram.
4. Since it is very speed analysis technique, more products can be analyzed with existing resources.
5. Operational cost is Low when compared to HPLC.
6.  Less solvent consumption.
7.  Uses of fine particle (2μm) for packing of stationary phase make analysis fast.

Disadvantages of UPLC:

1. In UPLC analysis the main disadvantage occurs are life of columns, during analysis high pressure developed because the particle size. Increase pressure reduces the life of the columns. Due to increased pressure requires more maintenance and reduces the life of the columns of these types. Using stationary phase of particle size 2μm perform better analysis without the adverse effects of high pressure.

Applications:

1. Now a days in pharmaceutical industry the demand of UPLC instrument is very high, because of the high resolution of peaks and short run time makes the work less with reliable cost.
2. We can generate more accurate data with UPLC.
3. The UPLC/MS system is used to generate a data which solved the complexity of the compound. By using MS as a detector with UPLC the interpretation of analysis is go to depth. Such analysis is very use full in bio-analytical field.
4. UPLC is used in the testing of Herbal products.
5. UPLC used to analyse, amino acids, natural medicines, herbal actives and drug content in Human plasma.