MA 334 / ANALYSIS OF ALPROSTADIL
BY HPLC WITH POST-COLUMN DERIVATIZATION
Since Prostaglandin E1 has low UV absorbance, the analysis of the formulations could be difficult. The addition of post-column derivatization increases the sensitivity and specificity of the analysis.
METHOD
Analytical Conditions
Analytical Column: Reversed-phase C18 column (150 x 4.6 mm)
Or alternative conditions:
Mobile Phase: 25% Acetonitrile – 75% of 0.0067 M Potassium Phosphate buffer pH 6.3
Post-column Conditions
Post-Column System: Pinnacle PCX
Calibration
Alprostadil: 0.1 ug/mL – 10 ug/mL β-Naphthol (Internal Standard): 0.25 ug/mL – 10 ug/mL Make stock solutions of Alprostadil and β-Naphthol in anhydrous Ethanol. Prepare working standards by appropriate dilution of stock solution with mobile phase.
Alprostadil (Prostaglandin E1) is a drug that has vasodilation properties and is used to treat erectile dysfunction (ED) and other medical conditions. This medication is available in injectable form and in suppository form.
Temperature: 37 °C
Flow Rate: 1 mL/min
Mobile Phase: 30% Acetonitrile – 70% of 0.02 M Potassium Phosphate monobasic (adjusted to pH 3)
Injection Volume: 20 uL
Reactor Volume: 2 mL
Reactor Temperature: 60 °C
Reagent: 1 mol/L Potassium Hydroxide
Reagent Flow Rate: 1 mL/min
Detection: UV 278 nm
