Determination of tacrolimus in rabbit aqueous humor by liquid chromatography-electrospray ionization tandem mass spectrometry.
A simple, sensitive and selective liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of tacrolimus (FK506) in rabbit aqueous humor. After a simple protein-precipitation by methanol, the post-treatment samples were separated on a reversed-phase, Thermo-Hypersil-BDS-C18 column with a mobile phase of a mixture of 0.1% formic acid in water, methanol and acetonitrile (5:85:10, v/v/v). Tacrolimus and ritonavir (internal standard, IS) were all detected by the selected reaction-monitoring (SRM) mode. The method developed was validated in rabbit aqueous humor with a daily working range of 0.5-100 ng/ml with correlation coefficient, r>0.99 and a sensitivity of 0.5 ng/ml as lower limit of quantification, respectively. This method was fully validated for the accuracy, precision, possible matrix effect and stability. The method proved to be accurate and specific, and was applied to the pharmacokinetic study of tacrolimus in rabbit aqueous humor.
Yuan
Jin
J
Zhongshan Ophthalmic Center, Sun Yat-sen University, The State Key Laboratory of Ophthalmology, Guangzhou, Guangdong Province, China.
Chen
Jia-Qi
JQ
Xie
Zhi-Yong
ZY
Zhai
Jia-Jie
JJ
Zhou
Shi-You
SY
D000818:Animals; D001082:Aqueous Humor / Q000737:chemistry*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007166:Immunosuppressive Agents / Q000032:analysis* / Q000493:pharmacokinetics; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D016559:Tacrolimus / Q000032:analysis* / Q000493:pharmacokinetics
D007166:Immunosuppressive Agents; D016559:Tacrolimus