If you are a decision maker in the business of science, a scientist who may request such a technique, are a technician doing this technique, or are simply interested, then this report will provide you a simple introduction to the area of HPLC.HPLC stands for high performance liquid chromatography. The first record of chromatography – as we know the practice now – was in the early 1900s, when a botanist wanted to separate pigments from a plant. Back then, gravity and some liquid solutions were used to pull apart the pigments to its constituent components.
Chromatography has Morphed through the years and is now a highly regulated, technical ability, requiring instrumentation with startup prices beginning at 30-50 million dollars, trained technicians to carry out the analysis, troubleshoot instrumentation maintenance and collapse, and compute and interpret results. In the U.S., what is hplc equipment is dominated by the Waters and Agilent Corporation, although Shimadtzu Corporation HPLC instrumentation is a fantastic option for some applications.
Both Waters and Agilent components are incompatible with one another; therefore, once a company decides to purchase a single, that provider is heldhostage to deal exclusively with that brand. Additionally, since the applications required for Waters and Agilent data evaluation is proprietary, external support can be expensive, as competition is sparse, and the exceptional software interface unlike any other product in existence.
With the right Instrumentation and method, modern HPLC analysis may be run in 2-3 minutes. More realistically, many methods require a run time of 30 minutes for each sample, and 30 minutes to get a clean or control, and 30 minutes for every standard. Together with prep time for each sample, standard, and management – as well as data analysis, documentation and communication results – total analysis time could be prohibitive. That said the Following are a few basic requirements for HPLC analysis.
HPLC is always performed in the liquid state. HPLC analysis can be carried out only if the molecule of interest remains in solution. If the molecule of interest cannot go into alternative – or it precipitates from solution – then analysis would not be accurate, or, a precipitate with clog HPLC tubing, costing tens of thousands of dollars to repair. Liquid solvents are utilized to execute the analysis. The liquid solvents are known as the mobile phase. Normal cases of aqueous mobile phase may consist of acetonitrile, methanol and water – with or without pH adjustment and/or buffer. A normal non-polar mobile phase solvent is hexane. Since human health dangers exist with a number of the mobile phase solvents used, it is essential that procedures be in place to protect everyone in the building and in the area, in the event of an incident.