Ion Exchange Chromatography (IEC) <ul><li>Elution by means of salt of pH gradient </li></ul><ul><li>For high polar compounds </li></ul>Time Ionic strength
IEC coupled to Mass Spectrometry: SACI No discharge phenomena (low voltage source) Few application because of analyte degradation and ion suppression phenomena Never applied till now ESI & APCI Discharge phenomena for the high voltages in API sources
Aim: IEC Analysis of M3G and M6G Optimal source: APCI, ESI or SACI? Morphine 3-glucoronide (M3G) Morphine 6-glucoronide (M6G)
M3G and M6G separation with a linear gradient 5-100% B A : H 2 O B : H 2 O + 0.5% formic ac. + 10mM ammonium acetate SACI ESI Cation Exchange Chromatography (CEC) Injection of 500pg Chromatographic analysis:
Intensitiy (counts/s) Results : SACI ESI Signal suppression (in source current ~ 40000nA) Good signal (in source current ~ 2 4nA) 2 4 6 8 10 12 Time [min] 0 200 400 600 800 Intensitiy (counts/s) 2 4 6 8 10 12 Time [min] 0.0 0.5 1.0 1.5 2.0 5 x10 10.11 8.42 Morphine-6- Glucuronide Morphine-3- Glucuronide
Conclusions: <ul><li>SACI is the only source suitable to coupling ion exchange chromatography with mass spectrometry </li></ul><ul><li>Analysis in urine is performed by SACI without any sample pre-tratment tanks to a low matrix effect </li></ul><ul><li>Good separation of analytes obtained with CEC </li></ul>
Future steps: Next experiments with potential and salt concentration variation Application of LC-CEC-SACI-MS for screening of other addictive drug metabolites in biological sample.
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