Highly sensitive assay for the determination of therapeutic peptide desmopressin in human plasma by UPLC-MS/MS

• Main Authors: Shiva Kumar Gudlawar (Author), Nageswara Rao Pilli (Author), Sridhar Siddiraju (Author), Jaya Dwivedi (Author)
• Format: Book
• Published: Elsevier, 2017-06-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.
• Notes: 2095-1779; 10.1016/j.jpha.2013.11.002
• Keywords: Desmopressin; Solid-phase extraction (SPE); Ultra performance liquid chromatography-tandem mass spectrometry; Method validation; Pharmacokinetics; Therapeutics. Pharmacology; RM1-950
• Source: Journal of Pharmaceutical Analysis, Vol 7, Iss 3, Pp 196-202 (2017)

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 d8) was used as an internal standard. Analyte and the internal standard were extracted from 200 µL of human plasma via 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 (r2≥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.