Hi !

Here you have to develop and optimize a method for detecting lysophospholipids in a finished liposomal formulation (non-pharmacopoeial), by employing reversed-phase HPLC with UV or ELSD detection, you can use a C18 column and gradient elution (typically with acetonitrile–water containing ammonium formate or formic acid). Optimize the parameters for retention, resolution, and peak shape using known standards. Validate the method per ICH Q2(R2) guidelines for specificity, linearity, accuracy, precision, LOD, and LOQ. Also you have to ensure sample prep includes liposome disruption (e.g., with ethanol or Triton X-100) for efficient lysophospholipid extraction.

Reference in support:

Article Simultaneous determination of lysophospholipids by high-perf...

Thank you, Mani Sharma

The most reliable approach for detecting lysophospholipid (LPL) impurities in finished liposomal products involves a combination of chromatographic separation and mass spectrometric detection. High-performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD) or charged aerosol detection (CAD) provides sensitive quantification without requiring chromophores, while mass spectrometry (LC-MS/MS) offers superior specificity for structural confirmation. Sample preparation typically involves organic solvent extraction (chloroform:methanol mixtures) to isolate lipids while maintaining vesicle integrity, followed by normal-phase chromatography using silica columns with gradient elution (e.g., chloroform/methanol/ammonium hydroxide) to separate LPLs from intact phospholipids. For enhanced sensitivity in trace analysis, derivatization with fluorescent tags like naphthalene-2,3-dicarboxaldehyde (NDA) prior to HPLC with fluorescence detection can achieve detection limits below 0.1%. Orthogonal validation using 31P-NMR spectroscopy is recommended for absolute quantification, as it directly detects the phosphorus headgroup with minimal sample preparation. Critical method validation parameters should include specificity against matrix components, detection limits below typical specification thresholds (often

Hi

To determine phospholipids, we used thin-layer chromatography and HPLC with ELSD-detector. We determined the amount of phospholipid impurities and their quantitative criteria in stability studies and degradation studies. We developed control methods ourselves depending on the composition of the drug.