Pharmaceutical analysis and Quality control
Date of submission: 20 – 02 - 07
APPLICATIONS OF HPLC
1.The wide verity of packing materials allows the separation of
most chemical species.Chemical Separations
can be accomplished using HPLC by
utilizing the fact that certain compounds have different
migration rates given a particular column and mobile phase.
Thus, the chromatographer can separate compounds (more on
chiral separations) from each other using HPLC; the extent or
degree of separation is mostly determined by the choice of
stationary phase and mobile phase.
e.g:a) Separation of Isomers of Amino Salicylic Acid
b) Separation of Isomers of Aminobenzoic Acid
2.The different types of detectors available permit the sensitive
detection of most chemical types, accurately and precisely.
3.Micro particulate packing materials give excellent separation of
4.The short column (3-10 cm) in routine use allow fast
5. A combination of HPLC and spectrometric techniques allows
almost simultaneous quantification and identification of solutes
of compounds or drugs by HPLC is a crucial part of
any HPLC assay. In order to identify any compound by HPLC a
detector must first be selected. Once the detector is selected and
is set to optimal detection settings, a separation assay must be
developed. The parameters of this assay should be such that a
clean peak of the known sample is observed from the
chromatograph. The identifying peak should have a reasonable
retention time and should be well separated from extraneous
peaks at the detection levels which the assay will be performed.
To alter the retention time of a compound, several parameters
can be manipulated. The first is the choice of column, another is
the choice of mobile phase, and last is the choice in flow rate7.Purity check:
refers to the process of separating or extracting the
target compound from other (possibly structurally related)
compounds or contaminants. Each compound should have a
characteristic peak under certain chromatographic conditions.
Depending on what needs to be separated and how closely related
the samples are, the chromatographer may choose the conditions,
such as the proper mobile phase, to allow adequate separation in
order to collect or extract the desired compound as it elutes from
the stationary phase. The migration of the compounds and
contaminants through the column need to differ enough so that
the pure desired compound can be collected or extracted without
incurring any other undesired compound.
8. Tablet dissolution of pharmaceutical dosages are also
9. Shelf-life determinations of pharmaceutical products can be
10. Pharmaceutical quality control and separation of many
e.g: a)Separation of Albuterol and Amoxicillin
b) Separation of Creatine, Creatinine and Histidine
c) Separation of Guaifenesin and Terbutaline
d) Separation of Epinephrine and Epinephrine Sulfonate
11.Separation of Active Compounds in Drug Formulation
12.Analysis of antibiotics can also be done by HPLC.
13. Detection of endogenous neuropeptides in brain extracellular
14. Stability of aspartame in the presence of glucose and
15.HPLC also identify anabolic steroids in serum, urine, sweat,
16.Determination of cocaine and metabolites in meconium.
of compounds by HPLC is the process of
determining the unknown concentration of a compound in a
known solution. It involves injecting a series of known
concentrations of the standard compound solution onto the HPLC
18. Quantification of DEET in Human Urine.
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