An analytical method based on ultra-performance liquid chromatography with positive ion electrospray ionization(ESI) coupled with tandem mass spectrometry(UPLC–MS/MS) was developed and validated for the determination...An analytical method based on ultra-performance liquid chromatography with positive ion electrospray ionization(ESI) coupled with tandem mass spectrometry(UPLC–MS/MS) was developed and validated for the determination of therapeutic peptide desmopressin in human plasma. A desmopressin stable labeled isotope(desmopressin d_8) was used as an internal standard. Analyte and the internal standard were extracted from200 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 m M ammonium formate buffer as the mobile phase. The calibration curve obtained was linear(r^2≥0.99)over the concentration range of 1.01–200 pg/m L. 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.展开更多
文摘An analytical method based on ultra-performance liquid chromatography with positive ion electrospray ionization(ESI) coupled with tandem mass spectrometry(UPLC–MS/MS) was developed and validated for the determination of therapeutic peptide desmopressin in human plasma. A desmopressin stable labeled isotope(desmopressin d_8) was used as an internal standard. Analyte and the internal standard were extracted from200 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 m M ammonium formate buffer as the mobile phase. The calibration curve obtained was linear(r^2≥0.99)over the concentration range of 1.01–200 pg/m L. 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.
