Quantitative determination of sirolimus in dog blood using liquid chromatography-tandem mass spectrometry, and its applications to pharmacokinetic studies.
A rapid, sensitive method of detecting sirolimus in blood was developed and applied in pharmacokinetic studies employing deionized water for hemolysis and a weakly basic mobile phase to enhance chromatographic peak intensity. Dog blood samples were processed via liquid-liquid extraction and the amounts of sirolimus and tacrolimus, an internal standard, were quantified by LC-MS/MS. Specificity, the lower limit of quantification, linearity, accuracy, precision, dilution, recovery, matrix effects, robustness and stability were within the acceptable range for assay validation. The concentration of sirolimus was quantifiable in blood samples for up to 36 h after the dog had received a 3 mg/kg dose of sirolimus. These observations suggest that sirolimus can be detected at low levels in dog blood using a basic mobile phase and metal-free hemolysis. This method is therefore applicable to pharmacokinetic studies in dogs.
Lee
Jong-Hwa
JH
Research and Development Division, Korea Institute of Toxicology, Daejeon, Republic of Korea.
Cha
Kwang-Ho
KH
Cho
Wonkyung
W
Park
Junsung
J
Park
Hee Jun
HJ
Cho
Youngseok
Y
Hwang
Sung-Joo
SJ
D000818:Animals; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D004285:Dogs; D004355:Drug Stability; D007166:Immunosuppressive Agents / Q000097:blood*; D057230:Limit of Detection; D020123:Sirolimus / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D053719:Tandem Mass Spectrometry / Q000379:methods*
D007166:Immunosuppressive Agents; D020123:Sirolimus