Application:
Pharmaceutical field
Environment study
Forensic study
Clinical application
Food and flavor
Therapeutically drug monitoring
Biochemistry
Industrial field
Cosmetic
Quality control
Diagnostic study
Qualitative analysis
Quantitative analysis
Pharmaceutical field:
Tablet dissolution of pharmaceutical dosages.
Shelf life determination of pharmaceutical products.
Identification of counterfeit drug product
Pharmaceutical quality control.
Cosmetic
The HPLC procedure is suitable for confirmation of the presence of preservatives in
cosmetic products like skin care product etc.
Environmental study:
Phenol in drinking water.
Identification of diphenhydramine in sediment samples.
Biomonitoring o PAH pollution in high-altitude mountain lakes through the analysis
of fish bite.
Estrogens in coastal waters. The sewage sources.
Toxicity of tetracycline degradations products to environmentally bacteria.
Assessment of TNT toxicity in sediment.
Forensic study:
A mobile HPLC apparatus at dance particle-on-site identification and
quantification of drug Ecstasy.
Identification of anabolic steroids in serum, urine, sweat and hair.
Forensic analysis of textile dyes.
Determination of cocaine and metabolites in meconium.
Simultaneous quantification of psychotherapeutic drugs in human plasma.
Food and flavor:
Ensuring soft drink consistency and qualify.
Analysis of vicinal diketone in beer.
Sugar analysis in fruit juices.
Polycyclic aromatic hydrocarbons in Brazilian vegetables and fruits.
Trace analysis of military high explosives in agricultural crops.
Stability of aspartame in the presence of glucose and vanillin.
Therapeutically drug monitoring:
This technique is widely used in application ranging from monitoring,
identifying and characterizing drug impurities to investigating deficit mass
balance, drug product discoloration, and drug product counterfeiting. It is use for
the analysis of antibiotics, sedatives , steroids and analysis.
Biochemistry
HPLC found one of its earliest application in the separation of nucleosides and in
part was responsible for the rapid development in the understanding of nucleic
and biochemistry .
Subsequently, chromatography has found a wide application n the determination
of nucleic acid composition in elucidating nucleic acid metabolism and also in
quantitating nucleotide pools.
Industrial field:
HPLC has it own importance because of its sensitivity and efficiency.
It is used for the analysis of condensed aromatics, surfactants, propellants and dyes.
It is used to determine the saturated contents of heavy petroleum products.
It is used for the analyzing complex mixtures, purifying chemical compound, developing
processes for synthesizing chemical compounds, isolating natural products that have
biological characteristics and predicting physical properties ( e.g molecular weight,
distribution of polymers).
Quality control:
HPLC is used to ensuring purify of raw material.
It is used in process testing to control and improve process yields.
It is used for quantitative assay of final product to assure Conformance to
specification.
It is used for evaluating product stability and monitoring degradation.
Diagnostic study:
Analysis their precursors and metabolites can provide diagnosis of diseases such
as Parkinson’s disease, heart disease and muscular dystrophy.
However, given how physiologically wide-spread these molecules are, their
analysis and subsequent conclusions about patient health must be done carefully.
HPLC has ability to separate and compare molecules to a higher magnitude than
other techniques, making it a great candidate for such diagnostic purpose.
Qualitative analysis:
HPLC is used to identify the some pure compound in sample. A small quantity of
the pure sample of the substance, which is supposed to be present in the
analytical sample is added to the sample. If the assumption about the presence of
that compound was correct the height of the peak due to that compound will rise
and if the assumption was wrong, a new peak will appear on the chromatogram.
The components separated are generally identified by mass spectrometry or infra
red spectroscopy.
Quantitative analysis:
Quantitative analysis is accomplished by measuring the peak heights and peak areas and
comparing these with a calibration curve.
Different known amounts of a substances are mixed with the carrier gas and mixture
passed through the chromatographic height us drawn. This is the calibration curve.
If several compounds are present in the analytical mixture , a separate calibration curve is
drawn for each compound. The peak height is length of perpendicular drawn from the top
off the peak to the base line.
The concentration also be calculated by measuring the area under the peak forming curve.
This is called the peak area. The peak area is found by multiplying the peak height by half
width of the peak ( width measured at the middle of the peak height.
