title: Comprehensive quantitative analysis of purines and pyrimidines in the human malaria parasite using ion-pairing ultra-performance liquid chromatography-mass spectrometry.
abstract: Targeted metabolite profiling has aided in the understanding of a variety of biological processes in the malaria parasite as well as in drug discovery. A fast and sensitive analytical method, based on ion-pairing reversed phase ultra-high performance liquid chromatography tandem mass spectrometry (IP-RP-UPLC-MS/MS), was optimized for the simultaneous analysis of intracellular levels of 35 purine and pyrimidine nucleobases, nucleosides, and nucleotides. This analytical method allows for chromatographic separation of highly polar metabolites using reverse phase chemistry within 15 min. The analytical performance of the method was evaluated and successfully applied to the quantification of purines and pyrimidines in Plasmodium falciparum and its host cell, the human erythrocyte. In addition, this method can be customized to include other related metabolites such as NADPH and NADP, among others.
authors.affiliation: Department of Biochemistry and Virginia Tech Center for Drug Discovery, Blacksburg, VA 24061, United States.
authors.forename: Christian D
authors.initials: CD
authors.lastname: Laourdakis
authors.affiliation: Department of Biochemistry and Virginia Tech Center for Drug Discovery, Blacksburg, VA 24061, United States.
authors.forename: Emilio F
authors.initials: EF
authors.lastname: Merino
authors.affiliation: Department of Food Science and Technology, Virginia Tech, Blacksburg, VA 24061, United States.
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Neilson
authors.affiliation: Department of Biochemistry and Virginia Tech Center for Drug Discovery, Blacksburg, VA 24061, United States. Electronic address: bcassera@vt.edu.
authors.forename: Maria B
authors.initials: MB
authors.lastname: Cassera
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004912:Erythrocytes / Q000737:chemistry / Q000469:parasitology; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D013058:Mass Spectrometry / Q000379:methods*; D010963:Plasmodium falciparum / Q000737:chemistry*; D011687:Purines / Q000032:analysis* / Q000737:chemistry; D011743:Pyrimidines / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results
chemical_list: D011687:Purines; D011743:Pyrimidines
keywords: Human erythrocytes; Malaria; Plasmodium falciparum; Purines and pyrimidines; UPLC–MS
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