Abstract 5494: A liquid chromatography-tandem mass spectrometric method for quantification of curcuminoids in cell culture medium and mouse plasma and their pharmacokinetics in mice

Abstract

Abstract Curcumin and tetrahydrocurcumin (THC) have been found as potent DNMT1 inhibitors, but they suffer from low oral bioavailability and rapid metabolism in vivo. To circumvent these problems, two curcumin analogs: 1,7-bis (3,4-dimethoxyphenyl)-4,4-dimethyl-1,6-heptadiene-3,5-dione (TMC) and 1,7-bis(3,4-dimethoxyphenyl)-4-cyclohexyl-1,6-heptadiene-3,5-dione (DMCHC) have been synthesized to enhance their stability by blocking the two metabolic sites, the phenolic and C4 methylene moieties. Both compounds have shown inhibitory activity on M. SssI similar to that of curcumin and THC (Poster, M1114, AAPS, 2009). Preclinical pharmacokinetics has yet to be performed. Purpose: A sensitive and rapid LC-MS/MS method for simultaneous determination of curcumin and its metabolite THC and analogs TMC and DMCHC in RPMI cell culture medium and mouse plasma has been developed and validated and their stability in these matrices and their pharmacokinetics in mice were evaluated. Method: A triple quadrupole mass spectrometer with electro-spray (ESI) ionization was used for quantification. Spiked curcuminoids and the internal standard, hesperetin were extracted from mouse plasma with ethyl acetate, and the components were separated on a BetaBasic C8 column. The ion transitions at m/z 369→177, m/z 373→137, m/z 425→191, m/z 465→191 and m/z 303→153 for curcumin. THC, TMC, DMHC, and hesperetin, respectively, were monitored for their quantification. These curcuminoids were incubated in mouse plasma or cell culture at different temperatures and aliquots were removed at different times up to 24 hrs. Cassette dose of curcumin and TMC (10 mg/kg each) was intraperitoneally administered to CD1F2 mice and timed plasma samples were collected up to 8 hr. Curcuminoids in these samples were quantified using this method. Result: The lower limit of quantifications for all four curcuminoids was found to be 1 ng/mL in both mouse plasma and cell culture medium. The within-day coefficients of variation were found to be below 15% and the accuracy was in the range of 85% to 115%. Curcuminoids degraded in two phases with terminal half lives of 186, 813, 724, and 2000 min for curcumin, IHC, TMC, and DMCHC, respectively, in cell culture medium. In plasma, their respective half lives were 111, 232, 1202 and 3000 min. These data demonstrated that their stability are in the order of curcumin<THC<TMC< DMCHC in both matrices. Following an i.p. cassette dose, TMC showed more rapid absorption than curcumin (15 vs 60 min) and yielded a higher Cmax (0.30 vs 0.21 μM). Conclusion: A sensitive LC-MS/MS method for simultaneous quantification of these curcumionids has been established. TMC and DMCHC were more stable than curcumin in cell culture medium and mouse plasma. Further pharmacokinetic study is ongoing. Supported by NIH/NCI R21 CA135478, and BioMedical Mass Spectrometric Laboratory. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5494.