title: Validated LC-MS/MS method for simultaneous determination of doxorubicin and curcumin in polymeric micelles in subcellular compartments of MCF-7/Adr cells by protein precipitation-ultrasonic breaking method.
abstract: A specific and sensitive LC-MS/MS with protein precipitation- ultrasonic breaking method has been developed and validated for simultaneous determination of doxorubicin (DOX) and curcumin (Cur) in DOX and Cur co-loaded hyaluronic acid-vitamin E succinatemicelles [(DOX + Cur)-polymeric micelles (PMs)] in subcellular compartments of resistant MCF-7/Adr cells. Sequential extraction of four subcellular protein fractions (cytosolic, membrane/organelle, nucleic and cytoskeleton) was performed directly from MCF-7/Adr cells after incubation with (DOX + Cur)-PMs. An ultrasonic breaking-methanol precipitation method was used for extraction of the fractions, and the micelle breaking efficiency with methanol was 98.1 and 97.6% for DOX and Cur, respectively. The analytes were analyzed using positive electrospray ionization coupled with multiple reaction monitoring. The calibration curves were linear over a concentration range of 0.5-400 ng/mL for DOX and 2-2000 ng/mL for Cur, and the recovery for the two analytes were >85% with negligible matrix effect. The intra-day and inter-day precision was <10.80% and relative error was within ±7.70%. The developed method was successfully applied for subcellular determination of DOX and Cur in MCF-7/Adr cells. Moreover, Cur and (DOX + Cur)-PMs had a marked promoting effect on the distribution of DOX in the nucleic protein fraction.
authors.affiliation: Modern Research Center for Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, People's Republic of China.
authors.forename: Jinling
authors.initials: J
authors.lastname: Wang
authors.affiliation: Modern Research Center for Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, People's Republic of China.|School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.affiliation: Modern Research Center for Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, People's Republic of China.
authors.forename: Wenzhuan
authors.initials: W
authors.lastname: Ma
authors.affiliation: Modern Research Center for Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, People's Republic of China.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Wang
authors.affiliation: Modern Research Center for Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, People's Republic of China.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Tu
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D003474:Curcumin / Q000032:analysis*; D004317:Doxorubicin / Q000032:analysis*; D006801:Humans; D061986:MCF-7 Cells; D008823:Micelles; D011108:Polymers / Q000737:chemistry*; D013347:Subcellular Fractions / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014465:Ultrasonics
chemical_list: D008823:Micelles; D011108:Polymers; D004317:Doxorubicin; D003474:Curcumin
keywords: HA-VES co-delivery micelles; LC-MS/MS; curcumin; doxorubicin; protein precipitation-ultrasonic breaking method; subcellular distribution