US EPA

3858248 

Journal Article 

Quantitative determination of dobutamine in newborn pig plasma samples by HPLC-MS/MS 

Albóniga, OE; Alonso, ML; Blanco, ME; González, O; Grisaleña, A; Campanero, MA; Alonso, RM 

2017 

Yes 

Journal of Pharmaceutical and Biomedical Analysis
ISSN: 0731-7085
EISSN: 1873-264X 

Elsevier B.V. 

145 

178-185 

English 

28666164 

10.1016/j.jpba.2017.06.050 

WOS:000410872200023 

A novel gradient reverse phase high performance liquid chromatography tandem mass spectrometry (HPLC/MS-MS) was performed as a method for the determination of dobutamine hydrochloride (DOB) in newborn pig plasma samples. It was developed and validated after optimization of sample treatment and various chromatographic and mass spectrometric conditions. Trimethoxydobutamine (TMD) was used as internal standard. Heptafluorobutyric acid (HFBA) and ethyl acetate were used for the treatment of plasma samples. The separation of dobutamine and internal standard was done using a Kinetex F5 (50×2.1mm, 2.6μm, 100Å) analytical column. The mobile phase was a mixture of acetonitrile and HCOOH 0.01%. The column oven temperature was optimized at 40° C and the flow rate was 0.25mL/min. DOB and TMD were detected by multiple reaction monitoring (MRM) mode in ESI+, using a cone voltage (CV) of 25V and a collision energy (CE) of 25eV. The weighted calibration curve (1/x(2)) was found to be linear over the concentration range of 1-100ng/mL (r(2)>0.999). The limit of quantification (LLOQ) of the method was 1ng/mL. The values of selectivity, carryover, LLOQ, linearity, accuracy, precision, matrix effect, stability and recovery obtained meet the acceptable range according to European Medicines Agency (EMA) and Food and Drug Administration (FDA) guidelines. The method was efficiently applied to quantify DOB in plasma samples from a pharmacokinetic/pharmacodynamic study in a disease model of newborn piglet. 

Dobutamine; HPLC–MS/MS; Newborn pig; Plasma