Exenatide is the first approved glucagon-like peptide 1 receptor agonist subcutaneously or intramuscularly injected for the treatment of type 2 diabetes mellitus. Typical therapeutic plasma concentrations are in the low pg/mL range, therefore requiring ultra-sensitive quantification. To enable the accurate evaluation of pharmacokinetic studies, we established a UPLC-MS/MS assay with a lower limit of quantification (LLOQ) of 5 pg/mL (1.2 pM) using 200 μL of plasma, validated according to FDA’s and EMA’s pertinent guidelines. Exenatide was isolated from plasma with solid phase extraction utilizing anion-exchange sorbent. Quantification was performed with positive electrospray ionization tandem mass spectrometry in the selected reaction monitoring mode. The calibrated concentration range of 5–10,000 pg/mL was linear showing correlation coefficients >0.99. Interday and intraday accuracy ranged from 97.5% to 105.4% with corresponding precision of <10.9%. Accuracy at the LLOQ ranged from 93.0% to 102.5% with corresponding precision of <15.9%. Because of the validity of a 10-fold dilution QC (accuracy 111.2%), the assay is suitable for exenatide quantification up to 100,000 pg/mL. The ultra-sensitive assay’s applicability was demonstrated by the quantification of exenatide plasma concentrations and pharmacokinetics after intravenous and nasal administration to beagle dogs.

艾塞那肽是第一种经皮下或肌内注射批准的胰高血糖素样肽1受体激动剂，用于治疗2型糖尿病。典型的治疗性血浆浓度在低pg / mL范围内，因此需要超灵敏的定量。为了能够准确评估药代动力学研究，我们建立了UPLC-MS / MS分析法，使用200μL血浆的定量下限（LLOQ）为5 pg / mL（1.2 pM），并根据FDA和EMA的相关准则进行了验证。通过使用阴离子交换吸附剂的固相萃取从血浆中分离出艾塞那肽。在选定的反应监测模式下，采用正电喷雾电离串联质谱进行定量。校准浓度范围为5-10，000 pg / mL是线性的，显示相关系数> 0.99。日间和日内准确性在97.5％至105.4％的范围内，相应的准确性<10.9％。LLOQ的准确度范围为93.0％至102.5％，相应的准确度为<15.9％。由于10倍稀释QC（准确度111.2％）的有效性，该测定适用于高达100,000 pg / mL的艾塞那肽定量。在对比格犬静脉和鼻内给药后，艾塞那肽血浆浓度和药代动力学的定量证明了超灵敏测定法的适用性。2％），该测定适用于高达100,000 pg / mL的艾塞那肽定量。在对比格犬静脉和鼻内给药后，艾塞那肽血浆浓度和药代动力学的定量证明了超灵敏测定法的适用性。2％），该测定适用于高达100,000 pg / mL的艾塞那肽定量。在对比格犬静脉和鼻内给药后，艾塞那肽血浆浓度和药代动力学的定量证明了超灵敏测定法的适用性。