Nature of the main contaminant in the drug primaquine diphosphate: SFC and SFC–MS methods of analysis

• Published in: Journal of pharmaceutical and biomedical analysis 2007-02, Vol.43 (3), p.937-944
• Main Authors: Brondz, Ilia, Ekeberg, Dag, Bell, David S., Annino, Amy R., Hustad, Jan Arild, Svendsen, Robert, Vlachos, Vaso, Oakley, Paul, Langley, G. John, Mohini, Thite, Amaury, Cazenave-Gassiot, Mikhalitsyn, Felix
• Format: Article
• Language: English
• Subjects: Aminoquinolines - analysis; Anti-malaria drug; Antimalarials - analysis; Chiral chromatography; Chromatography, High Pressure Liquid; Chromatography, Supercritical Fluid; Mass Spectrometry; Positional isomers; Primaquine; Primaquine - analysis; Quinocide; SFC–MS; Spectrometry, Mass, Electrospray Ionization; Spectrophotometry, Ultraviolet; Stereoisomerism
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2006.09.017

The drug primaquine diphosphate is used for causative treatment of malaria. Using HPLC–MS and GC–MS, this research group was previously able to show that the main contaminant of primaquine is the positional isomer quinocide [I. Brondz, D. Mantzilas, U. Klein, D. Ekeberg, E. Hvattum, M.N. Lebedeva, F.S. Mikhailitsyn, G.D. Soulimanov, J. Roe, J. Chromatogr. B: Anal. Technol. Biomed. Life Sci. 800 (2004) 211–223; I. Brondz, U. Klein, D. Ekeberg, D. Mantzilas, E. Hvattum, H. Schultz, F. S. Mikhailitsyn, Asian J. Chem. 17 (2005) 1678–1688]. Primaquine and quinocide are highly toxic substances which can have a number of side effects upon use in medical treatment. A standard for quinocide is not typically commercially available. In the present work, supercritical fluid chromatography–mass spectrometry (SFC–MS) with two different columns was used to achieve a shorter analysis time for the separation between the positional isomers quinocide and primaquine in primaquine diphosphate and to elucidate additional information about differences in their MS fragmentation. Unlike using HPLC–MS, it was possible to achieve the differential fragmentation of positional isomers at branching points using the SFC–MS technique. The desired short analysis time was achieved using SFC equipped with a Discovery HS F5 column and the differential fragmentation of positional isomers during SFC–MS provides information on the differences in the structure of these substances. Using a Chiralpak AD-H chiral column, it was possible to resolve the enantiomers in primaquine and separate quinocide from those enantiomers.