N desmethylclozapine

N-Desmethylclozapine, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), MPP⁺, and Leu methyl ester (LME) were purchased from Sigma-Aldrich (St. Louis, MO). Clozapine and clozapine N-oxide were obtained from the NIMH Chemical Synthesis and Drug Supply Program (Research Triangle Institute, RTP NC). Lipopolysaccharide (LPS strain O111:B4) was purchased from Calbiochem (San Diego, CA). WST-1 was purchased from Dojindo Laboratories (Gaithersburg, MD). Cell culture ingredients were obtained from Invitrogen (Carlsbad, CA). [³H]DA was purchased from PerkinElmer Life Sciences (Boston, MA). The polyclonal anti-tyrosine hydroxylase (TH) antibody was purchased from CHEMICON International (Temecula, CA). The polyclonal ionized calcium-binding adaptor molecule 1 (Iba-1) antibody was purchased from Wako Chemicals USA (Richmond, VA). The monoclonal CD11b antibody was purchased from AbDSerotec (Raleigh, NC). The biotinylated secondary antibodies were purchased from Vector Laboratories (Burlingame, CA).

Chlorpromazine, clozapine, olanzapine, and quetiapine were purchased from Cerilliant (Round Rock, TX, USA). DesmethylChlorpromazine and 7-hydroxy-Chlorpromazine were acquired from TRC Canada (Toronto, ON, Canada). N-desmethylclozapine was obtained from Sigma-Aldrich (St. Louis, MO, USA). N-desmethylolanzapine was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Norquetiapine was acquired from Biovision (Milpitas, CA, USA). The internal standards Chlorpromazine-d3 and quetiapine-d8 were obtained from Cerilliant (Round Rock, TX, USA). Butyl methacrylate (BMA) (99%), ethyleneglicol dimetacrylate (EGDMA, 98%), 1-propanol (HPLC grade), vinyltrimethoxysilane (VTMS) 1,4-butendiol (99%), and 2,2-azobisisobutylnitrile (AIBN) were purchased from Sigma–Aldrich (St. Louis, MO, USA). Fused silica capillary (530 μm i.d. × 10 cm) was acquired from NST (São Paulo, Brazil). Acetonitrile, methanol (HPLC grade), ammonium acetate, and formic acid were supplied by JTBaker (Phillisburg, NJ, USA). Hydrochloric acid and sodium hydroxide were purchased from Sigma–Aldrich (St. Louis, MO, USA). Aqueous solutions were prepared with ultrapure water from a Milli-Q, Millipore system (18.2 MΩ cm) (São Paulo, SP, Brazil).

The chemicals and reagents, including Altanserin, antipyrine, atenolol, buspirone, cimetidine, citalopram, N-desmethylclozapine, diphenhydramine, doxepin, fluoxetine, gabapentin, indomethacin, metoclopramide, propranolol, risperidone, and Way-100635 were purchased from Sigma Aldrich (St. Louise, Missouri, US). The calibration curves and matrices for the in vitro equilibrium brain-to-plasma distribution experiments, were prepared using BSA purchased from Sigma Aldrich (≥ 98%, St. Louise, Missouri, US, Product no.: A7906 Lot. No.: SLCH8448) and brain homogenate from BioIVT (Westbury, NY, US, Product no. RAT00BRAINMZA, lot no. RAT415237). For the in vitro binding experiments, BSA (purity > 98%) was obtained from Beijing SeaskyBio Technology Co. Ltd. (Beijing, North China, China. Product no.: BSAS, Lot.: 541), and Sprague–Dawley brain homogenate was purchased from BioIVT (Westbury, NY, US, Product no. RAT00BRAINMZA, Lot no.: RAT 472873) and Shanghai Biotechnology Co. Ltd (Shanghai, East China, China. Lot.: 20,221,213). Other chemicals and solvents were of analytical grade and obtained from a commercial supplier.