Gudlawar Shiva Kumar, Pilli Nageswara Rao, Siddiraju Sridhar, Dwivedi Jaya
Department of Chemistry, Banasthali Vidyapeeth, Jaipur 304022, India.
Wellquest Clinical Research Laboratories, Ramanthapur, Hyderabad 500013, India.
J Pharm Anal. 2017 Jun;7(3):196-202. doi: 10.1016/j.jpha.2013.11.002. Epub 2013 Dec 27.
An analytical method based on ultra-performance liquid chromatography with positive ion electrospray ionization (ESI) coupled with tandem mass spectrometry detection (UPLC-MS/MS) was developed and validated for the determination of therapeutic peptide desmopressin in human plasma. A desmopressin stable labeled isotope (desmopressin d) was used as an internal standard. Analyte and the internal standard were extracted from 200 µL of human plasma solid-phase extraction technique using Oasis WCX cartridges. The chromatographic separation was achieved on an Aquity UPLC HSS T3 column by using a gradient mixture of methanol and 1 mM ammonium formate buffer as the mobile phase. The calibration curve obtained was linear ( ≥0.99) over the concentration range of 1.01-200 pg/mL. Method validation was performed as per FDA guidelines and the results met the acceptance criteria. The results of the intra- and inter-day precision and accuracy studies were well within the acceptable limits. The proposed method was successfully applied to pharmacokinetic studies in humans.
建立了一种基于超高效液相色谱-正离子电喷雾电离（ESI）串联质谱检测（UPLC-MS/MS）的分析方法，并对其进行了验证，用于测定人血浆中的治疗性肽去氨加压素。使用去氨加压素稳定同位素标记物（去氨加压素d）作为内标。采用Oasis WCX柱，通过固相萃取技术从200 μL人血浆中提取分析物和内标。在Acquity UPLC HSS T3柱上进行色谱分离，以甲醇和1 mM甲酸铵缓冲液的梯度混合物作为流动相。在1.01-200 pg/mL的浓度范围内获得的校准曲线呈线性（r≥0.99）。按照FDA指南进行方法验证，结果符合验收标准。日内和日间精密度及准确度研究结果均在可接受范围内。该方法成功应用于人体药代动力学研究。
