Hplc (high performance liquid chromatography) is one of the sophisticated analytical tools in the present science. It is given prominent importance due to its attributes like
- High sensitivity i.e. ability to evaluate samples of very minute concentrations like in nano-gram and picogram
- Detect precisely the closely similar molecules and also
- Highest accuracy in the identification of components of most complex mixtures.
This is possible in hplc analysis due to efficient separation of molecules under pressure over a large surface area and also the availability of highly sensitive detectors.
But unlike other analytical techniques, HPLC analysis is not easy and trouble shooting is very important to run the test smoothly. So one is advised to go for requires prior hplc training.
Application of hplc chromatography is vast and it finds its use in
1. Clinical diagnosis of diseases, disorders.
2. In scientific research for analysis.
3. In pharmaceutical and food industry for quality control.
[alert-success]HPLC analysis in clinical diagnosis: [/alert-success]
Many disorders related to body metabolism, those related to endocrine and exocrine gland secretion, alteration in body fluids are diagnosed by HPLC analysis of concerned fluids.
For example estimation of metabolites of purines, pyrimidines or other metabolites from plasma, cerebrospinal fluid and urine samples in patients.
Estimation of corticoids from plasma in disorders of adrenal gland which secretes an endocrine hormone.
[alert-success]HPLC application in scientific research:[/alert-success]
HPLC system is a mandatory tool in most of the labs involved in medical, biological, chemical, biochemical, phytochemical i.e plant chemical research.
It finds it application to analyse and quantify molecules, components of with chemistry and properties. Due to the principle of separation in HPLC similar molecules get separated and hence their detection, identification and quantification becomes easier.
[alert-success]HPLC applications in pharmaceutical industry: [/alert-success]
In pharmaceutical industry, hplc analysis is prominently used in quality control and also research and development plants.
- In quality control, it is used to check if the prepared or manufactured products are in compliance with the specified standards as per the pharmacopoeia and other drug regulating bodies. For those in quality control referring pharmacopoeia will give an idea of how the peak of the drug in the formulation should look, what solvents are suitable as mobile phases etc..
For this the formulation like injections, solutions, dissolved form of solid dosage forms are injected into HPLC to record the peaks of the individual constituents.
- In research and development wing, the formulations under development or any new chemical molecules are analysed to see their concentration in the blood after certain intervals of administration. This helps to evaluate the metabolic profile, plasma concentration, bio-availability etc of the formulation or chemical moieties under development.
- In plant constituents, there are many molecules with similarity in chemistry like isoflavones, glycosides, saponins etc. but different activity or nutritional value. These compound can’t be precisely determined by other methods, hence they are determined by HPLC analysis by means of separation into individual components and there by identification.
The effectiveness is further enhanced due to coupling of hplc detectors with MS, NMR etc.