Development and Validation of an HPLC-RI Method for the Simultaneous Determination of DPPC, Palmitic Acid, and Cholesterol in Bovine Pulmonary Surfactant

IF 1.8 4区医学 Q4 BIOCHEMICAL RESEARCH METHODS

Biomedical Chromatography Pub Date : 2025-06-19 DOI:10.1002/bmc.70151

Mahsa Aghaei, Mahjoobeh Ehsani, Malihe Keramati

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引用次数: 0

Abstract

This study aimed to develop and validate an HPLC-RI method for the simultaneous determination of dipalmitoylphosphatidylcholine (DPPC), palmitic acid, and cholesterol in bovine pulmonary surfactant, a critical treatment for infants suffering from Respiratory Distress Syndrome (RDS). The Iranian surfactant, Beraksurf, was used as the sample. A simple extraction procedure involving methanol dissolution and protein precipitation was employed, followed by HPLC analysis. The method underwent comprehensive validation for various parameters, including system suitability, repeatability (intra-assay precision), intermediate precision, linearity, accuracy, limit of detection (LOD), limit of quantification (LOQ), and robustness. The results demonstrated excellent system suitability. Repeatability and intermediate precision were confirmed with low %RSD values, while linearity was established with correlation coefficients (R²) exceeding 0.99 for all analytes. The method exhibited high accuracy, with recoveries ranging from 97% to 100%, and low LOD and LOQ values, indicating its sensitivity. Robustness testing revealed that minor variations in method parameters did not significantly impact results. This validated RP-HPLC-RID method provides a reliable and efficient approach for the analysis of key surfactant components, contributing to improved quality control in surfactant preparations and enhancing therapeutic strategies for managing RDS in infants.

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HPLC-RI同时测定牛肺表面活性剂中DPPC、棕榈酸和胆固醇的方法的建立和验证

本研究旨在建立并验证HPLC-RI同时测定牛肺表面活性剂中双棕榈酰磷脂酰胆碱（DPPC）、棕榈酸和胆固醇的方法，这是婴儿呼吸窘迫综合征（RDS）的关键治疗方法。以伊朗表面活性剂Beraksurf为样品。采用甲醇溶解和蛋白质沉淀的简单提取方法，然后进行高效液相色谱分析。该方法对系统适用性、可重复性（测定内精密度）、中间精密度、线性度、准确度、检出限（LOD）、定量限（LOQ）和鲁棒性等参数进行了综合验证。结果表明，该系统具有良好的适用性。重复性和中间精密度均具有较低的RSD值，线性关系良好，相关系数（R2）均大于0.99。该方法准确度高，回收率为97% ~ 100%，LOD和LOQ值低，灵敏度高。稳健性检验显示，方法参数的微小变化对结果没有显著影响。经验证的RP-HPLC-RID方法为表面活性剂关键成分的分析提供了一种可靠、高效的方法，有助于改善表面活性剂制剂的质量控制，提高婴儿RDS的治疗策略。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biomedical Chromatography 生物-分析化学

CiteScore

3.60

自引率

5.60%

发文量

268

审稿时长

2.3 months

期刊介绍： Biomedical Chromatography is devoted to the publication of original papers on the applications of chromatography and allied techniques in the biological and medical sciences. Research papers and review articles cover the methods and techniques relevant to the separation, identification and determination of substances in biochemistry, biotechnology, molecular biology, cell biology, clinical chemistry, pharmacology and related disciplines. These include the analysis of body fluids, cells and tissues, purification of biologically important compounds, pharmaco-kinetics and sequencing methods using HPLC, GC, HPLC-MS, TLC, paper chromatography, affinity chromatography, gel filtration, electrophoresis and related techniques.