Saci And Ion Exchange Chromatography
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Transcript of Saci And Ion Exchange Chromatography
Ion exchange chromatography coupled to SACI for Ion exchange chromatography coupled to SACI for the analysis of high polar low molecular weight drugsthe analysis of high polar low molecular weight drugs
Sara Crotti, ISB srlSara Crotti, ISB srl
XIV Discussion group on Mass SpectrometryXIV Discussion group on Mass Spectrometry
Surface Activated Chemical Ionization
Ion Exchange Chromatography (IEC)Ion Exchange Chromatography (IEC)
TimeTime
Ionic
str
ength
Ionic
str
ength
Elution by means of salt of pH gradient
For high polar compounds
IEC coupled to Mass Spectrometry:IEC coupled to Mass Spectrometry:
ESI&APCI
Discharge phenomena for Discharge phenomena for the high voltages in API the high voltages in API
sourcessources
SACI No discharge phenomena (low No discharge phenomena (low
voltage source)voltage source)
Few application because of Few application because of analyte degradation and ion analyte degradation and ion
suppression phenomenasuppression phenomena
Never applied till nowNever applied till now
Aim: IEC Analysis of M3G and M6GAim: IEC Analysis of M3G and M6G
Optimal source: APCI, ESI or SACI? Optimal source: APCI, ESI or SACI?
Morphine 3-glucoronide Morphine 3-glucoronide (M3G)(M3G)
Morphine 6-glucoronide Morphine 6-glucoronide (M6G)(M6G)
Preliminary infusion of M3G 50ng/ml:Preliminary infusion of M3G 50ng/ml:
In source In source fragmentationfragmentation
MW 461 MW 285
0
2.0
3.0
4.0
5 x105
150 200 250 300 350 400 450 500 550 m/z
286
1.0
Morphine-3- Glucuronide in source fragment
Inte
nsit
y (
cou
nts
/s)
APCIAPCI
Inte
nsit
y (
cou
nts
/s)
462
3.0
4.0
5.0
6 x105
2.0
1.0
0
150 200 250 350 400 450 550 m/z300 500
[M+H]+
Source noise Source noise
Preliminary infusion of M3G 50ng/ml:Preliminary infusion of M3G 50ng/ml:
ESIESI
0.0
2.0
4.0
6.0
x105
150 200 250 300 350 400 450 500 550 m/z
462
286
8
Morphine-3- Glucuronide in source fragment
[M+H]+
Preliminary infusion of M3G 50ng/ml:Preliminary infusion of M3G 50ng/ml:In
ten
sit
y (
cou
nts
/s)
SACISACI
M3G and M6G separation with a linear gradient 5-M3G and M6G separation with a linear gradient 5-100% B100% B
AA: H: H22OO
BB: H: H22O + 0.5% formic ac. + O + 0.5% formic ac. + 10mM ammonium acetate10mM ammonium acetate
SACI
ESI
Cation Exchange Cation Exchange Chromatography Chromatography
(CEC)(CEC)
Injection of Injection of
500pg500pg
Chromatographic analysis:Chromatographic analysis:
2 4 6 8 10 12Time [min]
0
200
400
600
800
Inte
nsit
iy (
cou
nts
/s)
Inte
nsit
iy (
cou
nts
/s)
2 4 6 8 10 12Time [min]
0.0
0.5
1.0
1.5
2.05
x10 10.11
8.42
Morphine-6- Glucuronide
Morphine-3- Glucuronide
Results :Results :
SACI
ESI
Signal suppressionSignal suppression(in source current (in source current
~~40000nA) 40000nA)
Good signalGood signal(in source current (in source current
~~224nA) 4nA)
SACI urine sample analysis:SACI urine sample analysis:
Urine Urine samplesample
Dil. 1:5Dil. 1:5In HIn H2200
1010l injl inj
2 4 6 8 10 12
Time [min]
0.0
0.5
1.0
1.5
2.0x10510.15
8.48
M3G
M6G
Inte
nsit
iy
EIC m/z 462 m/z 286
Matrix effect:Matrix effect:
20 negative 20 negative
urine spiked urine spiked
vs vs
Standard Standard
solutionsolution
% Var=% Var=
(Au-As)/As*100(Au-As)/As*100
sample%Area variation urine vs Water
samples
1 -6
2 -8
3 -6
4 -1
5 -5
6 -7
7 -4
8 -4
9 -5
10 -5
11 -5
12 -5
13 -0,5
14 -2
15 -1
16 -1
17 -4
18 -3
19 -3
20 -2
Conclusions:Conclusions:
SACI is the only source suitable to coupling ion SACI is the only source suitable to coupling ion exchange chromatography with mass spectrometryexchange chromatography with mass spectrometry
Analysis in urine is performed by SACI without any Analysis in urine is performed by SACI without any sample pre-tratment tanks to a low matrix effectsample pre-tratment tanks to a low matrix effect
Good separation of analytes obtained with CECGood separation of analytes obtained with CEC
Future steps:Future steps:
Next experiments with potential and salt Next experiments with potential and salt concentration variation concentration variation
Application of LC-CEC-SACI-MS for screening of Application of LC-CEC-SACI-MS for screening of other addictive drug metabolites in biological other addictive drug metabolites in biological
sample. sample.
Where do you can find us ?Where do you can find us ?
Web site: http://www.isbiotechnology.comWeb site: http://www.isbiotechnology.com