Rabee Mahmoud, Said Ragab A M, Naguib Ibrahim A
Research and Development Department, Heliopolis University, Cairo, 11785, Egypt.
Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Al-Azhar University, Cairo, 11751, Egypt.
BMC Chem. 2024 Oct 9;18(1):195. doi: 10.1186/s13065-024-01300-5.
Ectoine (ECT) has recently gained considerable interest in the healthcare sector due to its promising therapeutic benefits in a variety of human disorders. This research aimed to quantify the ECT plasma level in rats by creating and optimizing a sensitive and validated UPLC-MS/MS method. Prior to analysis, ECT extraction from the plasma samples was conducted via a protein precipitation procedure, using hydroxyectoine as an internal standard (IS). A 1.7 μm UPLC C8 column (100 mm × 2.1 mm) was selected for the chromatographic separation, using a gradient mobile phase consisting of acetonitrile and 0.05% formic acid. The electrospray ionization mass spectrometry (ESI-MS) was used to detect ECT in the positive ion mode. To determine the specific precursor and the product ions of ECT, multiple reaction monitoring (MRM) methods were carried out. The selected ion pair of ECT was 143.1 > 97 and 159.1 > 113.13 for the IS. The ECT's linearity range in rat plasma was found to be 1-1000 ng/mL, with a recovery rate of 96.48-97.37%. Consistent with FDA guidelines for bio-analytical method validation, the suggested method was validated. The method was efficiently employed to quantify the studied drug in spiked rat plasma with good accuracy and precision with no significant matrix effects. Furthermore, it was effectively used to investigate the pharmacokinetic behavior of ECT in rats after a single oral dose of 30 mg/kg.
由于依克多因（ECT）在多种人类疾病中具有潜在的治疗益处，近年来它在医疗保健领域引起了广泛关注。本研究旨在通过建立和优化一种灵敏且经过验证的超高效液相色谱-串联质谱（UPLC-MS/MS）方法来定量大鼠血浆中的ECT水平。在分析之前，采用蛋白质沉淀法从血浆样品中提取ECT，使用羟基依克多因作为内标（IS）。选用1.7μm的UPLC C8柱（100mm×2.1mm）进行色谱分离，流动相采用由乙腈和0.05%甲酸组成的梯度洗脱。采用电喷雾电离质谱（ESI-MS）在正离子模式下检测ECT。为了确定ECT的特定母离子和子离子，进行了多反应监测（MRM）方法。ECT选定的离子对为143.1＞97，内标的离子对为159.1＞113.13。发现ECT在大鼠血浆中的线性范围为1-1000 ng/mL，回收率为96.48-97.37%。按照美国食品药品监督管理局（FDA）生物分析方法验证指南，对所建议的方法进行了验证。该方法有效地用于定量加标大鼠血浆中的研究药物，具有良好的准确度和精密度，且无明显基质效应。此外，它还有效地用于研究大鼠单次口服30mg/kg剂量后ECT的药代动力学行为。
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