Comparison of electrospray, atmospheric pressure chemical ionization, and atmospheric pressure photoionization in the identification of apomorphine, dobutamine, and entacapone phase II metabolites in biological samples.
The applicability of different ionization techniques, electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI), and a novel atmospheric pressure photoionization (APPI), were tested for the identification of the phase II metabolites of apomorphine, dobutamine, and entacapone in rat urine and in vitro incubation mixtures (rat hepatocytes and human liver microsomes). ESI proved to be the most suitable ionization method; it enabled detection of 22 conjugates, whereas APCI and APPI showed only 12 and 14 conjugates, respectively. Methyl conjugates were detected with all ionization methods. Glucuronide conjugates were ionized most efficiently with ESI. Only some of the glucuronides detected with ESI were detected with APCI and APPI. Sulfate conjugates were detected only with ESI. MS/MS experiments showed that the site of glucuronidation or sulfation could not be determined, since the primary cleavage was a loss of the conjugate group (glucuronic acid or SO3), and no site-characteristic product ions were formed. However, it may be possible to determine the site of methylation, since methylated products are more stable than glucuronides or sulfates. Furthermore, the loss of CH3 is not necessarily the primary cleavage, and site characteristic products may be formed. Identification and comparison of conjugates formed from the current model drugs were successfully analyzed in different biological specimens of common interest to biomedical research. A fairly good relation was obtained between the data from in vivo and in vitro models of drug metabolism.
Department of Pharmacy, University of Helsinki, Finland.
Helena
H
Keski-Hynnilä
Mika
M
Kurkela
Elvor
E
Elovaara
Laurence
L
Antonio
Jacques
J
Magdalou
Leena
L
Luukkanen
Jyrki
J
Taskinen
Risto
R
Kostiainen
D000818:Animals; D001058:Apomorphine / Q000032:analysis* / Q000652:urine; D001274:Atmospheric Pressure; D002396:Catechols / Q000032:analysis* / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004280:Dobutamine / Q000032:analysis* / Q000652:urine; D022781:Hepatocytes / Q000737:chemistry; D006801:Humans; D007477:Ions; D008297:Male; D008862:Microsomes, Liver / Q000737:chemistry; D015394:Molecular Structure; D009570:Nitriles; D051381:Rats; D017207:Rats, Sprague-Dawley; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D002396:Catechols; D007477:Ions; D009570:Nitriles; D004280:Dobutamine; C071192:entacapone; D001058:Apomorphine