A quantitative LC-MS/MS method for determination of thiazolidinedione mitoNEET ligand NL-1 in mouse serum suitable for pharmacokinetic studies.

A quantitative LC-MS/MS method for determination of thiazolidinedione mitoNEET ligand NL-1 in mouse serum suitable for pharmacokinetic studies.

Pedada Kiran K; Zhou Xiang; Jogiraju Harini; Carroll Richard T; Geldenhuys Werner J; Lin Li; Anderson David J

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences

2014

2014-01

Thiazolidinedione (TZD) compounds have shown promise as antidiabetic, antibiotics, antifungal and neuroprotective agents. The mitochondrial effect of a novel mitoNEET ligand, NL-1 {5-[(3,5-di-tert-butyl-4-hydroxyphenyl)methyl]-1,3-thiazolidine-2,4-dione}, and other TZD compounds, is a newly proposed mechanism for the neuroprotective action of these TZD compounds. In this work, a sensitive LC-MS/MS assay has been developed and validated for quantification of NL-1 in mouse serum. Sample preparation involved an acetonitrile protein precipitation procedure with addition of an internal standard NL-2 {5-[(4-hydroxy-3,5-dimethyl-phenyl)methyl]thiazolidine-2,4-dione}. LC-MS/MS analysis utilized a Columbus C-18 HPLC column (2mmx50mm, 5mum). Chromatography employed a multiple step gradient program that featured a steep linear gradient (25-95% in 0.5min) of 15muM ammonium acetate (additive for eliminating carry-over) in 2% methanol mixing with increasing proportions of 100% methanol. The HPLC was interfaced to a QTrap 5500 mass spectrometer (AB Sciex) equipped with an electrospray ionization source used in a negative ionization mode. Multiple reaction monitoring (MRM) of m/z 334–\textgreater263 for NL-1 and m/z 250–\textgreater179 for NL-2 was done. The method had a linear range of at least 1-100ng/mL in serum. The intra-assay and inter-assay percent coefficient of variation (%CV) were less than 4% and accuracies (%RE) ranged from -2.7% to 2.0%. The analytical procedure gave 96-115% absolute extraction recovery of NL-1. The relative matrix effect was measured and found to be insignificant. The analyte in serum was confirmed to be stable during storage and treatment. The method is suitable for pharmacokinetic (PK) studies of the parent drug NL-1 based on the preliminary serum results from dosed NL-1 mouse studies.

%CV; %RE; 1-methyl-4-phenyl-1; 2; 3; 3-thiazolidine-2; 4-dione; 5-[3; 5-[4-hydroxy-3; 5-di-tert-butyl-4-hydroxyphenyl)methyl]-1; 5-dimethyl-phenyl)methyl]thiazolidine-2; 6-tetrahydropyridine; Animals; Carry-over; Chromatography; High Pressure Liquid/*methods; internal standard; IS; LC-MS/MS; Limit of Detection; LLOQ; lower limit of quantification; Methanol; Mice; mitoNEET; Mouse serum; MPTP; MRM; multiple reaction monitoring; NL-1; NL-2; percent coefficient of variation; peroxisome proliferator activated receptor-gamma; pharmacokinetic; PK; PPAR-gamma; QC; quality control; relative error; relative matrix effect; RME; SD; standard deviation; Tandem Mass Spectrometry/*methods; thiazolidinedione; Thiazolidinedione (TZD); Thiazolidinediones/*blood; TZD

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141–146

945-946