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A Rapid Isocratic High-Performance Liquid Chromatography Method for Determination of Cholesterol and 1,2-dioleoyl-sn-glycero-3-phosphocholine in Liposome-Based Drug Formulations

A Rapid Isocratic High-Performance Liquid Chromatography Method for Determination of Cholesterol and 1,2-dioleoyl-sn-glycero-3-phosphocholine in Liposome-Based Drug Formulations

Ramsharan Singh, Monsurat Ajagbe, Shastri Bhamidipati, Zafeer Ahmad, Imran Ahmad

J Chromatogr A. 2005 May 6;1073(1-2):347-53.

PMID: 15909540

Abstract:

A high-performance liquid chromatography (HPLC) method for the determination of cholesterol and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) in liposome-based drug formulations has been developed. Liposome formulations of anticancer agents (viz., paclitaxel, docetaxel, 7-ethyl-10-hydroxycamptothecin (SN38), doxorubicin, mitoxantrone and an antisense oligodeoxyribonucleotide, etc.) were prepared. These formulations contain DOPC, cholesterol and other lipids, such as tetramyristoyl cardiolipin or 1,3-bis(1,2-bis-tetradecyloxy-propyl-3-dimethylethoxyammonium bromide)propan-2-ol [(R)-PCL-2] in product-specific ratios. A simple HPLC method that uses isocratic elution and UV detection has been developed for simultaneous quantification of cholesterol and DOPC components of the liposome formulations. The chromatographic separation of these components is achieved using a C8 analytical column with 50 mM ammonium phosphate buffer (pH 2.7)-methanol (15:85, v/v) as mobile phase. Both cholesterol and DOPC peaks are well resolved and free of interference from other excipients or degraded impurities in the formulation. The method has been found to be linear (r > 0.999) over a wide concentration range of both analytes. This method offers the advantage of simultaneous quantitation of cholesterol and DOPC in various liposome-based formulations without any preprocessing of the sample, and has quantitation limits of 0.5 and 10 microg/mL for cholesterol and DOPC, respectively.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP4235954	1,2-Dioleoyl-sn-glycero-3-phosphocholine		4235-95-4	Price

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