title: A quantitative LC-MS/MS method for determination of thiazolidinedione mitoNEET ligand NL-1 in mouse serum suitable for pharmacokinetic studies.
abstract: 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 (2mm×50mm, 5μm). Chromatography employed a multiple step gradient program that featured a steep linear gradient (25-95% in 0.5min) of 15μM 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→263 for NL-1 and m/z 250→179 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.
authors: Department of Chemistry, Cleveland State University, 2121 Euclid Avenue, Cleveland, OH 44115, United States.
authors: Kiran K
authors: KK
authors: Pedada
authors: Department of Chemistry, Cleveland State University, 2121 Euclid Avenue, Cleveland, OH 44115, United States.
authors: Xiang
authors: X
authors: Zhou
authors: Department of Chemistry, Cleveland State University, 2121 Euclid Avenue, Cleveland, OH 44115, United States.
authors: Harini
authors: H
authors: Jogiraju
authors: Department of Pharmaceutical Sciences, Northeast Ohio Medical University, 4209 State Route 44, Rootstown, OH 44272, United States.
authors: Richard T
authors: RT
authors: Carroll
authors: Department of Pharmaceutical Sciences, Northeast Ohio Medical University, 4209 State Route 44, Rootstown, OH 44272, United States.
authors: Werner J
authors: WJ
authors: Geldenhuys
authors: Department of Pharmaceutical Sciences, Northeast Ohio Medical University, 4209 State Route 44, Rootstown, OH 44272, United States.
authors: Li
authors: L
authors: Lin
authors: Department of Chemistry, Cleveland State University, 2121 Euclid Avenue, Cleveland, OH 44115, United States. Electronic address: d.anderson@csuohio.edu.
authors: David J
authors: DJ
authors: Anderson
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D057230:Limit of Detection; D000432:Methanol; D051379:Mice; D053719:Tandem Mass Spectrometry / Q000379:methods*; D045162:Thiazolidinediones / Q000097:blood*
chemical_list: D045162:Thiazolidinediones; C500090:thiazolidine-2,4-dione; D000432:Methanol
keywords: %CV; %RE; 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; 5-[3,5-di-tert-butyl-4-hydroxyphenyl)methyl]-1,3-thiazolidine-2,4-dione; 5-[4-hydroxy-3,5-dimethyl-phenyl)methyl]thiazolidine-2,4-dione; Carry-over; IS; LC–MS/MS; LLOQ; MPTP; MRM; Mouse serum; NL-1; NL-2; PK; PPAR-γ; QC; RME; SD; TZD; Thiazolidinedione (TZD); internal standard; lower limit of quantification; mitoNEET; multiple reaction monitoring; percent coefficient of variation; peroxisome proliferator activated receptor-gamma; pharmacokinetic; quality control; relative error; relative matrix effect; standard deviation; thiazolidinedione