HPLC Method development Training -Derex Labs - Hyderabad

I.Method Development:

a.Introduction: Purpose and Scope of analytical method development. Its Advantages and applications
b. Setup the method development goals
c.Understand the chemistry of analytes
i.pH
ii.Solubility
iii.Melting point
iv.Polarity
v.Basic structure
vi.Synthesis process
vii.Possible degradants/ Impurities
d.Literature collection or search for compendial methods
e.Select a suitable chromatography method like HPLC or GC
f.Started with Primary conditions
g.Optimization of
i.Column
1.Choice of column
2.Polarity
3.Dimensions
4.Reproducibility
5.Bad peak shape
6.temperature
ii.Mobile phase
1.Reverse Phase or Normal Phase
2.Gradient or Isocratic
3.Solvent Strength
4.Buffer Capacity
5.Buffer UV absorption
6.Preferred buffers
7.Effect of pH and Ion pairing
iii.Selecting or optimize Analyte concentration
iv.System suitability
1.Resolution
2.Peak Tailing
3.Column Theoretical plates and
4.Relative standard deviation
5.
v.Detection Sensitivity and Selectivity
1.Types of Detectors
2.General Considerations
3.Selecting suitable detector
4.Choice of Wave lengths
5.Optimize the S/N ratio
6.Optimize the maximum S/N for Assay and other peaks

vi.Completing the method development
1.Checking of repeatability
2.Analyzing available samples (Different lots)
3.Perform the preliminary validation
a.Precision
b.Linearity
c.Accuracy
d.Robustness

II.Trouble Shooting LC and GC Systems:
LC System: During the method development, these are the common problems arise
A.Pressure (High or Low)
B.Leaks
C.Retention time problems
D.Inaccurate Results
E.Base line noise instability
F.Poor Peak Shapes
G.Extra Peaks
H.Injector problems
GC System:
A.No Peaks
B.Poor sensitivity with normal retention
C.Negative peaks
D.Base line drift
E.Constant base line drift in one direction
F.High background signal

III.Prevention of Problems:
A.Selecting initial Components
B.Routine preventive maintenance
C.Early replacement of critical parts
D.Record Keeping

For Details & Registration contact Ms.Jyothsna on 9949974890/9246882890 . Visit us at www.derexlabs.com