Pharmaceutics & Pharmacokinetics Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India; Academy of Scientific and Innovative Research, New Delhi, 110025, India.
Pharmaceutics & Pharmacokinetics Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India.
J Pharm Biomed Anal. 2020 Feb 20;180:113074. doi: 10.1016/j.jpba.2019.113074. Epub 2019 Dec 23.
PSTi8 is a 21 amino acid pancreastatin inhibitory peptide that demonstrated potent antidiabetic activity in insulin resistant rodent models. The goal of the current work is to establish and validate the LC-ESI-MS/MS bioanalytical assay of PSTi8 in mice plasma in order to unveil its pharmacokinetic (PK) behaviour for the first time. The MS detection of PSTi8 and diprotin A (internal standard, IS) was conducted with Q1/Q3 SRM transitions at 607.80 ([M+4 H])/771.20 and 342.20/229.10, respectively using positive ESI. Phenomenex Aqua 5μ 125A (250 × 4.6 mm) column was utilized to separate PSTi8 and IS with a mobile phase consists of MeOH-0.1 % formic acid (1:1, v/v) using 0.4 mL/min flow rate. SPE using medium anion exchange cartridge (Oasis MAX) was used for the extraction of analyte and IS from the mice plasma and the extraction recovery was found to be >55 %. PSTi8 displayed good linearity across the 5-1000 ng/mL concentrations range. The intra- and inter- day accuracy was observed between 99.44-110.20 % and 99.66-110.93 %, respectively. The intra- and inter- day precision was observed between 2.61-4.03 % and 2.90-7.16 %, respectively. The intra-day and inter-day accuracy and precision data was within the 100 ± 15 % nominal values recommended by the United States Food and Drug Administration bioanalytical guidance. The LC-MS/MS assay was validated effectively to investigate the PSTi8 plasma concentrations following intravenous and intraperitoneal PK studies in mice. The absolute bioavailability of PSTi8 was 52.74 ± 13.50 %.
PSTi8 是一种 21 个氨基酸的胰高血糖素抑制肽，在胰岛素抵抗的啮齿动物模型中表现出强大的抗糖尿病活性。本研究的目的是建立和验证 PSTi8 在小鼠血浆中的 LC-ESI-MS/MS 生物分析测定法，以便首次揭示其药代动力学（PK）行为。使用正离子 ESI，通过 Q1/Q3 SRM 跃迁，在 607.80 ([M+4H])/771.20 和 342.20/229.10 处对 PSTi8 和二肽基肽酶 IV（内标，IS）进行 MS 检测。使用 Phenomenex Aqua 5μ 125A（250×4.6mm）柱，以甲醇-0.1%甲酸（1:1，v/v）为流动相，流速为 0.4mL/min，分离 PSTi8 和 IS。使用中等阴离子交换小柱（Oasis MAX）进行 SPE，从小鼠血浆中提取分析物和 IS，提取回收率>55%。PSTi8 在 5-1000ng/mL 浓度范围内具有良好的线性。日内和日间准确度分别为 99.44-110.20%和 99.66-110.93%。日内和日间精密度分别为 2.61-4.03%和 2.90-7.16%。日内和日间准确度和精密度数据均在 100±15%的美国食品和药物管理局生物分析指南推荐的名义值范围内。该 LC-MS/MS 测定法经过有效验证，可用于研究 PSTi8 在小鼠静脉和腹腔 PK 研究后的血浆浓度。PSTi8 的绝对生物利用度为 52.74±13.50%。
