A liquid chromatography–tandem mass spectrometric method for quantification of curcuminoids in cell medium and mouse plasma

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. In this paper, a simple liquid chromatography-tandem mass spectrometric method was developed for the determination of these four curcuminoids in cell medium and mouse plasma. The method showed linearity from 1 to 1000 ng/mL with the lower limit of quantification of 1 ng/ mL in cell medium, and 5 ng/mL in mouse plasma for all test curcuminoids. The within-day coefficients of variation were found to be below 15% and the accuracy was in the range of 85% to 115%. This method was subsequently used to evaluate their stability in these matrices and a pilot pharmacokinetics of curcumin, DMCHC and TMC in mice after an intraperitoneal (I.P.) cassette dosing of 10 mg/kg each. Curcuminoids degraded in two phases with terminal half lives of 186, 813, 724, and 2000 min for curcumin, THC, 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 is in the order curcumin<THC<TMC< DMCHC in both matrices. Following an i.p. cassette dose, Both TMC and DMCHC showed the prolonged elimination half life (1.0, 1.0 h respectively vs 0.4 h for curcumin) and an increased drug exposure as described by the area under the curve (0.64, 0.98 μM*h respectively vs 0.4 μM*h for curcumin). KeywordsCurcuminoids; Liquid Chromatography-Mass Spectrometry; Cell medium; mouse plasma; stability; pharmacokinetics * Corresponding Author: Zhongfa Liu, Ph.D., College of Pharmacy, the Ohio State University, 500 W. 12 th Ave. RM 151, Columbus, OH 43210, Tel: (614) Fax: (614) 292-7766, liu.550@osu.edu . Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. -4]. Due to its potential inhibition of mutagenesis and chemically induced carcinogenesis via perturbation of multiple molecular targets during these processes, it has become a vital lead molecule for the development of various potential anti-cancer agents that can be used for the prevention and/or...

A high-throughput quantification method of curcuminoids and curcumin metabolites in human plasma via high-performance liquid chromatography/tandem mass spectrometry

“…We have reported two LC-MS/MS methods for the quantitation of curcumin, THC, TMC, and DMCHC [9] and the quantitation of curcumin and COG [10]. Therefore, we first evaluated the same extraction and LC conditions for these curcuminoids and curcumin metabolites under the negative mode defined from Section 3.1.…”

“…Recently, we characterized the mass spectra of curcumin, its two synthetic analogs tetramethyl curcumin (TMC) and dimethylcyclohexyl curcumin (DMCHC), and its two metabolites tetrahydrocurcumin (THC) and curcumin O -glucuronide (COG) under a positive ion mode on a Thermo TSQ Quantum triple quadrupole mass spectrometer [9,10]. The protonated molecular ions are the predominant ions in their respective mass spectra; the major fragmentation pathway for curcumin, THC, TMC, and DMCHC is the cleavage of the heptenoid chain between the 3rd and the 4th carbon atoms (Scheme 1) [9]; and the major fragmentation for COG is glycosidic cleavage (Scheme 1) [10].…”

“…The protonated molecular ions are the predominant ions in their respective mass spectra; the major fragmentation pathway for curcumin, THC, TMC, and DMCHC is the cleavage of the heptenoid chain between the 3rd and the 4th carbon atoms (Scheme 1) [9]; and the major fragmentation for COG is glycosidic cleavage (Scheme 1) [10]. Two analytic methods for quantification of curcuminoids and COG [10] were established in both mouse and human plasma under a positive mode.…”

“…Two analytic methods for quantification of curcuminoids and COG [10] were established in both mouse and human plasma under a positive mode. These methods were used to characterize their stabilities and pharmacokinetics in mouse and human [9,10]. However, these two methods are incapable of measuring COS. To develop an integrative method for simultaneous quantification of curcumin, DMC, BDMC, COG, COS, and THC, the mass spectra and tandem mass spectra of these curcuminoids and curcumin metabolites under both positive and negative modes were compared.…”

Curcumin Differs from Tetrahydrocurcumin for Molecular Targets, Signaling Pathways and Cellular Responses

“…The degradation half lives of curcumin, and THC were 186 and 813 min respectively in cell culture medium. In plasma, their respective half-lives were 111 and 232 min [81]. It has been also reported that THC and curcumin increased Epigallocatechin-3-gallate uptake by greater than two-fold in MDCKII/MRP1 and HT-29 cells [82].…”