SAMPLE

Matrix: blood
Sample preparation: Condition a Bond-Elut C18 column with 2 volumes MeOH then 2
volumes water. Add 1 mL serum then 2 !L " n#$mL %roma&ine in MeOHrO. 1 M HCl
1'(8" to each column) wash with 2 volumes water) wash with 2 volumes .1 M acetic acid)
wash with MeOH$water) add 2 *+LL 1 mM ammonium acetate in MeOH) wait ,or ' s)
elute with vacuum) re%eat elution %rocess two more times. Combine eluates and eva%orate
them to dr-ness at ./-80 under com%ressed air. 1econstitute with 2 !L mobile
%hase) vorte2 1 s) in3ect ".-1 !L ali4uot. *MeOH$water was . mL MeOH(water /.(
'. %lus 2. !L concentrated HCl.5
HPLCVARIABLES
Column: 2. 6 7./ . 3im 8u%elco silica
Mobile phase: EtOH(MeC9(t-but-lamine :8(2(.. *Mi2 1 #allon EtOH with "" mL MeC9
and 1.: mL t-but-lamine.5
Flow rate:
In!e"tion #olume: ".-1
$ete"tor: %V 2.7
CHR&MA'&(RAM
Retention time: './) '.' *cis isomer5
Internal stan)ar): %roma&ine *..25
Limit o* )ete"tion: 2 n#$mL
&'HER S%BS'A+CES
Extra"te): amitri%t-line) desi%ramine) desmeth-ldo2e%in) imi%ramine) nortri%t-line)
%rotri%t-line
Simultaneous: 9-acet-l%rocainamide) %rocainamide) &imeldine) mor%hine) codeine)
tri,luo%era&ine) 2-h-dro2-desi%ramine) desmeth-lchlordia&e%o2ide) bu%rion) dia&e%am)
demo2e%am) chlordia&e%o2ide) %ro%o2-%hene) de2tro%ro%o2-%hene) cocaine) o2a%am)
trimi%ramine) mianserin) trime%ra&ine) lo2e%in) ,lu%hena&ine) methadone) tri,luo%roma&ine)
%henteramine) chlorimi%ramine
+oninter*erin,: thio%ro%a&ine
Inter*erin,: thiorida&ine) h-dro2-amo2a%ine) me%eridine) chlor%roma&ine) diso%-ramide)
am%hetamine) 2-h-dro2-imi%ramine) i%rindole) %-rilamine) %rometha&ine) %roli2in)
amo2a%ine
-E./&R$S
serum; normal %hase
REFERE+CE
Beierle)<.A.; Hubbard)1-=. Li4uid chromato#ra%hic se%aration o, antide%ressant dru#s( >.
?ric-clics) Ther.DrugMonit, 1:8') .) 2":-2:2.
SAMPLE
Matrix: blood
Sample preparation: 1 mL 8erum @ 1 mL 7. mM 9aOH @ . mL he2ane(iso%ro%anol :.(
.) shaAe ,or . min) centri,u#e. 1emove 7 mL o, the or#anic la-er and add it to . !L 2
mM HCl) shaAe ,or 2 min) centri,u#e. >n3ect a 2 !L ali4uot o, the a4ueous la-er.
HPLC VARIABLES
Column: 1 6 ' oct-l CB-tm-8%her C8 #lass column *Chrom%acA5
Mobile phase: MeC9(. mM 9aH2BO7 '.(/. ad3usted to %H 2.2 with %hos%horic acid
Flow rate: 012
In!e"tion #olume: 2
$ete"tor: %V 21
CHR&MA'&(RAM
Retention time: 313
Limit o* )ete"tion: 1 n#$mL
&'HER S%BS'A+CES
Simultaneous: nortri%t-line) amitri%t-line
-E. /&R$S
serum
REFERE+CE
Can Damme)M.; Molle)L.; Abi Ehalil)<. Fse,ul sam%le handlin#s ,or reversed %hase hi#h
%er,ormance
li4uid chromato#ra%h- in emer#enc- to2icolo#-) J.Toxicol.Clin.ToxicoL, 45267 23, .8:-/17.
SAMPLE
Matrix: blood
Sample preparation: Condition a 1 mL Anal-tichem c-ano%ro%-l 8BE cartrid#e with 1
mL water and 1 mL MeOH) do not allow to dr-. Add 1 mL serum @ 2. *2L .2 !#$mL
imi%ramine in . mM sodium n-he%tanesul,onic acid to the 8BE cartrid#e) wash with 1
mL water) 1 mL MeOH(water .(.) air dr- cartrid#e) elute with 1 mL MeOH(trieth-lamine
::.2(.8) eva%orate eluate to dr-ness under a stream o, nitro#en at 70) reconstitute
residue with 2. !L mobile %hase) in3ect a . !L ali4uot.
HPLC VARIABLES
Column: 1. 6 7./ .*2m 8%herisorb c-ano%ro%-l
Mobile phase: MeOH(2 mM %hos%horic acid containin# ..G 9)9-dieth-loct-lamine ..(
7.) %H was 2.7
Flow rate: 416
In!e"tion #olume: 60
$ete"tor: %V 217
CHR&MA'&(RAM
Retention time: 8
Internal stan)ar): imi%ramine *:5
Limit o* )ete"tion: 2 n#$mL
&'HER S%BS'A+CES
Simultaneous: metabolites
-E./&R$S
serum; 8BE
REFERE+CE
Emm)?.; LesAo)L.+.; BerAal)M.B. 8imultaneous determination o, do2e%in and nordo2e%in in
serum usin#
hi#h-%er,ormance li4uid chromato#ra%h-) J.Chromatogr., 45297 419, 77.-7.1.