title: Normal phase and reverse phase HPLC-UV-MS analysis of process impurities for rapamycin analog ABT-578: application to active pharmaceutical ingredient process development.
abstract: ABT-578, an active pharmaceutical ingredient (API), is a semi-synthetic tetrazole derivative of the fermented polyene macrolide rapamycin. Reverse phase (RP)-HPLC-UV-MS and normal phase (NP)-HPLC-UV-MS methods employing an LC/MSD trap with electrospray ionization (ESI) have been developed to track and map all significant impurities from the synthetic process. Trace-level tracking of key impurities occurring at various process points was achieved using complimentary methodologies, including a stability indicating reverse phase HPLC method capable of separating at least 25 starting materials and process-related impurities from the API (YMC-Pack Phenyl column, UV-MS, 210 nm) and a targeted reverse phase HPLC method capable of separating very polar compounds from crude reaction mixtures (Phenomenex Synergi Polar RP column, UV, 265 nm). In addition, a normal phase HPLC method condition with post-column modifier infusion is described for the separation of epimeric impurities, and analysis of aqueous-sensitive reactive species (YMC-Pack SIL column, UV-MS, 278 nm). Process control strategies were established with these combinations of analytical technologies for impurities analyses to enable a rich understanding of the ABT-578 process.
authors.affiliation: D-R45T, Process Analytical Chemistry, Global Pharmaceutical Research and Development, Abbott Laboratories, North Chicago, IL 60064-4000, USA. yong.chen@abbott.com
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.forename: Gregory M
authors.initials: GM
authors.lastname: Brill
authors.forename: Nancy J
authors.initials: NJ
authors.lastname: Benz
authors.forename: M Robert
authors.initials: MR
authors.lastname: Leanna
authors.forename: Madhup K
authors.initials: MK
authors.lastname: Dhaon
authors.forename: Michael
authors.initials: M
authors.lastname: Rasmussen
authors.forename: Casey Chun
authors.initials: CC
authors.lastname: Zhou
authors.forename: James A
authors.initials: JA
authors.lastname: Bruzek
authors.forename: John R
authors.initials: JR
authors.lastname: Bellettini
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004340:Drug Contamination; D015394:Molecular Structure; D004364:Pharmaceutical Preparations / Q000032:analysis / Q000737:chemistry; D015203:Reproducibility of Results; D020123:Sirolimus / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D013237:Stereoisomerism