Identification of adenine nucleotide-containing metabolites of bisphosphonate drugs using ion-pair liquid chromatography-electrospray mass spectrometry.
Bisphosphonates are synthetic pyrophosphate analogues, which are used as therapeutic drugs for the treatment of metabolic bone disorders. Some of these bisphosphonates can be metabolised in cells into non-hydrolysable nucleotide analogues. In this paper, we describe an ion-pairing high-performance liquid chromatography method that is compatible with negative ion electrospray mass spectrometry for the separation of these metabolites. Tandem mass spectrometry and collision-induced dissociation (CID) were used for identification of the metabolites. The CID mass spectra of bisphosphonate-adenine nucleotide adducts are very informative, because major fragment ions are formed by cleavage of the bisphosphonate moiety from the conjugate. The method was used for detection of the nucleotide metabolites of clodronate, tiludronate and etidronate in extracts from mammalian cells after treatment with bisphosphonates.
Auriola
S
S
Department of Pharmaceutical Chemistry, University of Kuopio, Finland.
Frith
J
J
Rogers
M J
MJ
Koivuniemi
A
A
Mönkkönen
J
J
D000227:Adenine Nucleotides / Q000032:analysis*; D000818:Animals; D001862:Bone Resorption / Q000517:prevention & control; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004002:Clodronic Acid / Q000032:analysis / Q000378:metabolism; D004164:Diphosphonates / Q000032:analysis* / Q000378:metabolism*; D012968:Etidronic Acid / Q000032:analysis / Q000378:metabolism; D008264:Macrophages / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice
D000227:Adenine Nucleotides; D004164:Diphosphonates; D004002:Clodronic Acid; C058651:tiludronic acid; D012968:Etidronic Acid