Ultra Pressure Liquid Chromatography

HPLC played a wide role in the laboratories in last 3 decades. HPLC is a dynamic and versatile in nature. It is dynamic in scope of application from capillary to preparative while versatile in the range of detection technique. In 1970s, HPLC was just introduced which have column packed irregularly with 10 micrometer size particles and pressure range was about 6500 psi pressure. In 1990, modification in particle size done which greatly decreased from 10 micrometer to 3-5 micrometer with the same pressure working at about 6500 psi pressure. But as time passes better accuracy and precision demanded from HPLC, in which sometime it fails to withstand with the assigned standards. Thus researches once again started to modify HPLC. To increase the efficiency 1st you have to decrease the particle size and second increase the pressure. Because smaller particle required higher pressure to force through the column.

And in 2004, Dr. J. Jorgensen from University of North Carolina and Dr. M. Lee from Brigham Young University published paper which give the introduction of Ultra pressure Liquid Chromatography.

They give the short information between HPLC and UPLC.

HPLC

• Broader peak width provides less resolution

• Less sample throughput comparatively.

• Sample injection volume is 20 micro liters.

• Pump operates at 2000-6000 psi pressure

• Particle size in stationary phase packing material is between 5-12 micrometers.

UPLC

• Smaller peak width provides better resolution and more number of peaks getting identified.

• Higher sample throughput with more information per sample.

• Sample injection volume is as less as 3-5 micro liters.

• Pump operates at 10,000 psi pressure

• Particle size in stationary phase packing material is less than 2 micrometer.

Applications

1. Drug Discovery

UPLC improves the drug discovery process by means of high throughput screening, combinational chemistry, high throughput in vitro screening to determine physiochemical and drug’s pharmacokinetics.

2. High throughput quantitative analysis

UPLC coupled with time of flight mass spectroscopy give the metabolic stability assay.

3. Analysis of Dosage form

It provides high speed, accuracy and reproducible results for isocratic and gradient analysis of drugs and their related substance. Thus method development time decrease.

4. Analysis of amino acids

UPLC used from accurate, reliable and reproducible analysis of amino acids in the areas of protein characterizations, cell culture monitoring and the nutritional   analysis of foods.

5. Determination of Pesticides

UPLC couples with triple Quadra-pole tandem mass spectroscopy will help in   identification of trace level of pesticides from water.

Thus Ultra Pressure Liquid Chromatography set a new standard in the science of chromatography. Working range with 15000 to 16000 psi pressure and column packed with less than 2 micrometer in size helped in various fields.