Singh Ramsharan, Ajagbe Monsurat, Bhamidipati Shastri, Ahmad Zafeer, Ahmad Imran
Process Development and Manufacturing Department, Research and Development, NeoPharm Inc., 1850 Lakeside Drive, Waukegan, IL 60085, USA.
J Chromatogr A. 2005 May 6;1073(1-2):347-53. doi: 10.1016/j.chroma.2004.12.036.
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.
已开发出一种用于测定基于脂质体的药物制剂中胆固醇和1,2 - 二油酰 - sn - 甘油 - 3 - 磷酸胆碱（DOPC）的高效液相色谱（HPLC）方法。制备了抗癌药物（即紫杉醇、多西他赛、7 - 乙基 - 10 - 羟基喜树碱（SN38）、阿霉素、米托蒽醌和一种反义寡脱氧核糖核苷酸等）的脂质体制剂。这些制剂含有DOPC、胆固醇和其他脂质，如四肉豆蔻酰心磷脂或1,3 - 双（1,2 - 双十四烷氧基 - 丙基 - 3 - 二甲基乙氧基溴化铵）丙 - 2 - 醇[（R） - PCL - 2]，其比例因产品而异。已开发出一种使用等度洗脱和紫外检测的简单HPLC方法，用于同时定量脂质体制剂中的胆固醇和DOPC成分。使用C8分析柱，以50 mM磷酸铵缓冲液（pH 2.7） - 甲醇（15:85，v/v）作为流动相，实现了这些成分的色谱分离。胆固醇和DOPC峰均得到良好分离，且不受制剂中其他辅料或降解杂质的干扰。已发现该方法在两种分析物的宽浓度范围内呈线性（r > 0.999）。该方法的优点是无需对样品进行任何预处理即可同时定量各种基于脂质体的制剂中的胆固醇和DOPC，胆固醇和DOPC的定量限分别为0.5和10 μg/mL。
