Validation of a high-performance liquid chromatography method for determining lysophosphatidylcholine content in bovine pulmonary surfactant medication

被引:0
作者
Aghaei, Mahsa [1 ,2 ]
Talari, Faezeh Shirgaei [1 ]
Mollahosseini, Afsaneh [3 ]
Keramati, Malihe [4 ]
机构
[1] ARC Bioassay, Iran Food & Drug Adm Accredited QC Lab Biopharmace, Tehran, Iran
[2] Univ Massachusetts Lowell, Dept Chem, One Univ Ave, Lowell, MA 01854 USA
[3] Iran Univ Sci & Technol, Dept Chem, Res Lab Spectrometry & Micro & Nano Extract, Tehran, Iran
[4] Pasteur Inst Iran, Dept Nanobiotechnol, New Technol Res Grp, Tehran, Iran
关键词
bovine pulmonary surfactant; HPLC validation; lysophosphatidylcholine (LPC); sample preparation; LUNG SURFACTANT; RESPIRATORY-DISTRESS; PHOSPHOLIPIDS; PROTEIN; HPLC; PHOSPHATIDYLCHOLINE; SEPARATION; PHASE; MS/MS; ACID;
D O I
10.1002/bmc.5926
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Pulmonary surfactant replacement therapy is a promising improvement in neonatal care for infants with respiratory distress syndrome. Lysophosphatidylcholine (LPC) is an undesirable component that can hinder surfactant proteins from enhancing the adsorption of surfactant lipids to balance surface tensions by creating a saturated coating on the interior of the lungs. A novel normal-phase liquid chromatography method utilizing UV detection and non-toxic solvents was developed and validated for the first time to analyze LPC in the complex matrix of pulmonary surfactant medication. The analytical method validation included evaluation of system suitability, repeatability, intermediate precision, linearity, accuracy, limit of detection (LOD), limit of quantification (LOQ), stability and robustness. The method yielded detection and quantification limits of 4.4 and 14.5 mu g/ml, respectively. The calibration curve was modified linearly within the LOQ to 1.44 mg/ml range, with a determination coefficient of 0.9999 for standards and 0.9997 for sample solutions. Given the lack of reliable published data on LPC analysis in pulmonary surfactant medications, this newly developed method demonstrates promising results and offers advantages of HPLC methodology, including simplicity, accuracy, specificity, sensitivity and an exceptionally low LOD and LOQ. These attributes contribute to considering this achievement as an innovative method.
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页数:11
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