A high-performance liquid-chromatographic assay for amphotericin B in a hydrophilic colloidal paste base

Area of Science:

• Analytical Chemistry
• Pharmaceutical Analysis

Background:

• Amphotericin B (AmB) is a critical antifungal agent.
• Pharmaceutical formulations containing AmB require robust analytical methods for quality control.
• Existing methods may not adequately address the complexity of multi-component pastes.

Purpose of the Study:

• To develop and validate a stability-indicating HPLC assay for amphotericin B (AmB) in a complex paste.
• To ensure accurate quantification of AmB in the presence of other active pharmaceutical ingredients and excipients.

Main Methods:

• Developed a stability-indicating high-performance liquid-chromatographic (HPLC) assay.
• Employed a liquid-liquid extraction technique using N,N-dimethylformamide (DMF) and cyclohexane to isolate AmB.
• Analyzed the DMF layer containing AmB using HPLC.

Main Results:

• Achieved a linear relationship between AmB concentration and peak area response (r = 0.9995) over the range of 5.0 x 10(-4) to 7.5 x 10(-3)% (w/v).
• Demonstrated high precision with a relative standard deviation of +/- 1.46% (n = 8).
• Reported an extraction efficiency of 100.6 +/- 2.4% (n = 5).

Conclusions:

• The developed HPLC method is suitable for the stability-indicating analysis of AmB in the specified paste formulation.
• The method provides accurate and precise quantification of AmB, supporting pharmaceutical quality control.
• The extraction procedure effectively isolates AmB from interfering components in the complex matrix.