title: LC-MS/MS method for determination of geldanamycin derivative GM-AMPL in rat plasma to support preclinical development.
abstract: A LC-MS/MS method for determining the concentration of the small molecule Hsp90 inhibitor, GM-AMPL, has been developed and validated in rat plasma to support preclinical development. 17-[2-(morpholine-4-yl)ethyl]amino-17-demethoxygeldanamycin (GM-AMPL) and the internal standard 17-allylamino-17-demethoxygeldanamycin (17-AAG) were sufficiently separated on a Venusil MP C18 column that was eluted with 80% methanol in water at 40°C. Quantification studies were performed with a multiple reaction monitoring (MRM) transition of m/z 657.3→614.3 and 584.3→541.3 for GM-AMPL and IS, respectively, in the negative ion mode. The present method exhibited good linearity (R>0.999) over the concentration range of 2-600ng/mL for GM-AMPL in rat plasma with a lower limit of quantification (LLOQ) of 2ng/mL. The intra-batch and inter-batch assay coefficients of variation (CV) were in range of 1.56-6.84% and 1.62-6.98%, respectively. The plasma samples were extracted with methanol to precipitate protein with extraction recovery in range of 84.09-95.25%. The matrix effect was determined as internal substance (IS) normalized matrix factor of 1.09, 1.18 and 1.05 for samples with three concentration levels of 4, 40 and 400ng/mL, respectively. This validated method was further applied to successfully determine the pharmacokinetic parameters and oral availability of GM-AMPL in Sprague-Dawley rats following intravenous injection and oral administration.
authors.affiliation: Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, 100050 Beijing, China.
authors.forename: Yan-ping
authors.initials: YP
authors.lastname: Li
authors.forename: Lin-yan
authors.initials: LY
authors.lastname: Gao
authors.forename: Kai-tong
authors.initials: KT
authors.lastname: Li
authors.forename: Shuai
authors.initials: S
authors.lastname: Meng
authors.forename: Jian-hua
authors.initials: JH
authors.lastname: Zhu
authors.forename: Dong
authors.initials: D
authors.lastname: Li
authors.forename: Jie
authors.initials: J
authors.lastname: Jin
authors.forename: Guang-zhi
authors.initials: GZ
authors.lastname: Shan
authors.forename: Zhuo-rong
authors.initials: ZR
authors.lastname: Li
mesh_terms: D000818:Animals; D016227:Benzoquinones / Q000737:chemistry* / Q000493:pharmacokinetics*; D002853:Chromatography, Liquid / Q000379:methods*; D005260:Female; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors; D047029:Lactams, Macrocyclic / Q000737:chemistry* / Q000493:pharmacokinetics*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D016227:Benzoquinones; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; C001277:geldanamycin