HPLC-MS-MS Determination of ZCZ-011, A Novel Pharmacological Tool for Investigation of the Cannabinoid Receptor in Mouse Brain Using Clean Screen FASt™ Column Extraction.
A high-performance liquid chromatography tandem mass spectrometry method was developed for the detection and quantification of 6-methyl-3-(2-nitro-1-(thiophen-2-yl)propyl)-2-phenyl-1H-indole (ZCZ-011) using 2-phenylindole as the internal standard (ISTD). ZCZ-011 was synthesized as a possible positive allosteric modulator with the CB1 cannabinoid receptor. The analytical method employs a rapid extraction technique using Clean Screen FASt™ columns with a Positive Pressure Manifold. FASt™ columns were originally developed for urine drug analysis but we have successfully adapted them to the extraction of brain tissue. Chromatographic separation was performed on a Restek Allure Biphenyl 5 µ, 100 × 3.2 mm column (Bellefonte, PA). The mobile phase consisted of 1:9 deionized water with 10 mmol ammonium acetate and 0.1% formic acid-methanol. The following transition ions (m/z) were monitored for ZCZ-011: 363 > 207 and 363 > 110 and for the ISTD: 194 > 165 and 194 > 89. The FASt™ columns lowered and stabilized the ion suppression over the linear range of the assay (40-4,000 ng/g). The method was evaluated for recovery, ion suppression, accuracy/bias, intraday and interday precision, bench-top stability, freeze-thaw and post-preparative stability. The method was successfully applied to brain tissue from C57BL/6J mice that received intraperitoneal (i.p.) injections with 40 mg/kg of ZCZ-011 or vehicle.
Department of Pharmacology and Toxicology, Virginia Commonwealth University, Box 980613, Richmond, VA 23298-0613, USA jlpoklis@vcu.edu.
Justin L
JL
Poklis
Department of Forensic Science, Virginia Commonwealth University, Richmond, VA, USA.
Deborah J
DJ
Clay
Department of Pharmacology and Toxicology, Virginia Commonwealth University, Box 980613, Richmond, VA 23298-0613, USA.
Bogna M
BM
Ignatowska-Jankowska
School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Aberdeen, UK.
Chiara
C
Zanato
Department of Pharmacology and Toxicology, University of Toronto, Toronto, ON, Canada.
Ruth A
RA
Ross
School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Aberdeen, UK.
Iain R
IR
Greig
Department of Pharmacology and Toxicology, Virginia Commonwealth University, Box 980613, Richmond, VA 23298-0613, USA.
Rehab A
RA
Abdullah
Department of Pharmacology and Toxicology, Virginia Commonwealth University, Box 980613, Richmond, VA 23298-0613, USA.
Mohammed A
MA
Mustafa
Department of Pharmacology and Toxicology, Virginia Commonwealth University, Box 980613, Richmond, VA 23298-0613, USA.
Aron H
AH
Lichtman
Department of Pharmacology and Toxicology, Virginia Commonwealth University, Box 980613, Richmond, VA 23298-0613, USA Department of Forensic Science, Virginia Commonwealth University, Richmond, VA, USA Department of Pathology, Virginia Commonwealth University, Richmond, VA, USA.
Alphonse
A
Poklis
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D007211:Indoles / Q000008:administration & dosage / Q000032:analysis* / Q000378:metabolism; D007274:Injections, Intraperitoneal; D057230:Limit of Detection; D016014:Linear Models; D051379:Mice; D008810:Mice, Inbred C57BL; D011485:Protein Binding; D043884:Receptor, Cannabinoid, CB1 / Q000302:isolation & purification* / Q000378:metabolism; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000592:standards; D013876:Thiophenes / Q000008:administration & dosage / Q000032:analysis*
C000604852:6-methyl-3-(2-nitro-1-(thiophen-2-yl)propyl)-2-phenyl-1H-indole; C578453:CNR1 protein, mouse; D007211:Indoles; D043884:Receptor, Cannabinoid, CB1; D013876:Thiophenes