EXPERIMENT 3

ASSAY OF ACETAMINOPHEN
TABLETS USING HPLC
QC2 LABORATORY
HPLC
• High Performance Liquid Chromatography
• Liquid Chromatography (LC)

• A separation technique based on a solid stationary phase and a liquid

mobile phase (USP 36).
HPLC
• STATIONARY PHASE

Separations are achieved by partition, adsorption, or ion-exchange

processes depending on the type of stationary phase used.

Most commonly used: silica, polymeric beads

HPLC
• CHROMATOGRAPHIC COLUMN

Includes stainless steel, lines stainless steel, and polymeric column,

packed with a stationary phase.
The length and inner diameter of the column affects the separation,
and therefore typical column dimensions are included in the individual
monographs.
HPLC
• MOBILE PHASE

A solvent or a mixture of solvents, as defined in the individual

monograph (USP 36).
HPLC
• APPARATUS

A liquid chromatogram consists of (USP 36):

• Reservoir containing the mobile phase
• Pump to force the mobile phase through the system at high pressure
• Injector to introduce the sample into the mobile phase
• Chromatographic column
• Detector
• Data collection device
HPLC
• GRADIENT ELUTION

The technique of continuously changing the solvent composition during

the chromatographic run.
Experiment 3
MOBILE PHASE

To prepare, measure 250 mL HPLC-grade methanol using a volumetric

flask. Transfer to 1000 mL volumetric flask and fill to volume.
Experiment 3
STANDARD SOLUTION

0.01 mg/mL of USP Acetaminophen RS in mobile phase

Experiment 3
SAMPLE SOLUTION

Weigh, and finely powder NLT 20 Tablets. Dissolve a portion of the

powder in Mobile phase to prepare a solution containing 0.01 mg/mL
of acetaminophen. To facilitate dissolution, shake the powder in Mobile
phase by mechanical means for 10 min, sonicate for 5 min, and dilute
with Mobile phase to volume. Pass a portion of this solution through a
filter of 0.5- μm or finer pore size, discarding the first 10 mL of the
filtrate. Use the clear filtrate.
Experiment 3
SAMPLE SOLUTION

Compute for the weight of powder containing ~100 mg of

acetaminophen
Computation