Page 77 - Instrumental Analysis - Pharm D Clinical- 07-PA403

Method development

1. Understanding the sample: matrix, approximate concentration
range and number of analytes

2. Knowing the physical and chemical properties of the analytes
including: pKa values, solubility, polarity.

3. Determination of the analysis requirements: Detection,
quantitation, characterization.

4. Selection of the analysis technique.

5. Determination of the initial conditions and Modulation of:

- stationary phase (composition, particle size, column length)

-mobile phase (composition, pH and effect of pH on analytes
ionization and retention)

- Elution mode selection

- Column temperature 63
Prof. Omayma Abdelrazak Amin

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Application of HPLC

HPLC is the most widely used technique for the
pharmacopeial

identification & quantification limit test of
of pharmaceutical compounds impurities

Prof. Omayma Abdelrazak Amin 64

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