State Key Laboratory of Drug Metabolism and Pharmacokinetics, Department of Pharmaceutical Sciences, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing 100850, China.
State Key Laboratory of Drug Metabolism and Pharmacokinetics, Department of Pharmaceutical Sciences, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing 100850, China; Guangxi Medical University, 22 Shuangyong Road, Nanning 530000, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Aug 15;1092:296-305. doi: 10.1016/j.jchromb.2018.05.042. Epub 2018 May 29.
HYD-PEP06 is a novel RGD-modified Endostar mimetic peptide with 30 amino acids that is intended to suppress the formation of neoplasm vessels. This assay was developed and validated to monitor the level of the peptide HYD-PEP06 in rat blood, using liquid chromatography tandem mass spectrometry (LC-MS/MS). HYD-PEP10, another peptide similar to the analyte, was used as an internal standard (IS). A triple quadrupole mass spectrometry in Multiple Reaction Monitoring (MRM) mode and an electrospray interface (ESI) in the positive mode were used for MS analysis. The analysis was optimized with addition of 0.3% formic acid (FA) into the mobile phase as well as with a needle washing solution to overcome the carryover effect. In addition, the carryover was reduced by optimizing the mobile phase gradient. Methanol was used as a diluent of working solutions to avoid any adsorption. Methanol:acetonitrile (1:1, v:v) containing 0.3% FA was employed to precipitate the blood samples. Unknown blood samples must be placed in ice bath immediately, and precipitating agents should be added within 30 min to ensure the stability of blood samples. The assay was established and validated. This method showed a good linear relationship for the HYD-PEP06 in the range of 10 ng·mL to 2000 ng·mL, with R > 0.99. HYD-PEP06 was determined with accuracy values (RE%) of -5.06%-8.54%, intra- and inter-day precisions (RSD%) of 3.13%-4.87% and 4.81%-9.42%. The method was successfully in monitoring the concentration of HYD-PEP06 in rat blood.
HYD-PEP06 是一种新型的 RGD 修饰的内皮抑素模拟肽，由 30 个氨基酸组成，旨在抑制新血管的形成。本实验采用液相色谱串联质谱法（LC-MS/MS）建立并验证了该方法，用于监测大鼠血液中 HYD-PEP06 肽的水平。HYD-PEP10 是一种与分析物类似的肽，被用作内标（IS）。三重四极杆质谱在多重反应监测（MRM）模式下，电喷雾接口（ESI）在正模式下用于 MS 分析。通过在流动相中加入 0.3%甲酸（FA）以及针洗涤溶液来优化分析，以克服残留效应。此外，通过优化流动相梯度来减少残留。甲醇用作工作溶液的稀释剂，以避免任何吸附。甲醇：乙腈（1:1，v:v）中含有 0.3%FA 用于沉淀血样。未知血样必须立即置于冰浴中，沉淀剂必须在 30 分钟内加入，以确保血样的稳定性。该方法已建立并验证。该方法在 10ng·mL 至 2000ng·mL 的范围内对 HYD-PEP06 显示出良好的线性关系，R > 0.99。HYD-PEP06 的准确度值（RE%）为-5.06%-8.54%，日内和日间精密度（RSD%）分别为 3.13%-4.87%和 4.81%-9.42%。该方法成功地监测了大鼠血液中 HYD-PEP06 的浓度。
