Ropinirole On Oasis MCX
Introduction
Ropinirole is a potent, basic drug with a low oral dose range. A bioanalytical method needs to be developed that can reach a lower limit of quantitation of 0.02 ng/mL, which is lower than what is typically required. The Oasis® MCX µElution 96-well plate was selected for its increased sensitivity through concentration, without drying down which allows reconstituting the extract.
Experimental
Test Conditions
|
Oasis MCX µ Elution 96-well plate: |
|
|
Condition: |
200 µL MeOH |
|
Equilibrate: |
200 µL H2O |
|
Load: |
400 µL diluted plasma sample (200 µL plasma diluted 1:1 with 4% H3P O4) |
|
Wash 1: |
200 µL 2% HCOOH |
|
Wash 2: |
200 µL MeOH |
|
Elute: |
2 X 25 µL 5% NH4OH in 90:10 MeOH:H2O (less MeOH used for a cleaner extract – less residual matrix components eluted.) |
|
Injection: |
8 µL (direct injection) |
|
Column: |
ACQUITY UPLC® BEH C18, 2.1 x 50 mm, 1.7 µm |
|
Mobile Phase A: |
10 mM NH4COOH, pH 9 |
|
Mobile P hase B: |
MeOH |
|
Flow Rate: |
0.5 mL/min |
|
Injection Volume: |
8.0 µL |
|
Column Temp: |
45 °C |
|
Sample Temp: |
15 °C |
|
Sample Diluent: |
25:75 H2O:MeOH |
|
Strong Needle Wash: |
60:40 ACN:IPA + 0.5% HCOOH (1200 µL) |
|
Weak Needle Wash: |
95:5 H2O:MeOH (500 µL) |
Gradient Table
|
Quattro Premier™ |
|
|
Desolvation Temp: |
350 °C |
|
Cone Gas Flow: |
50 L/Hr |
|
Desolvation Gas Flow: |
700 L/Hr |
|
Collision C ell P ressure: |
2.6 x 10 (-3) mbar |
|
MRM transitions monitored: |
Ropinirole m/z 261.2 > 113.95 ( ESI+) Citalopram (IS) m/z 325.2 > 108.85 (ESI+) For both analytes, the optimal cone voltage was 40 V. For Ropinirole and Citalopram, the collision energies were 18 eV and 25 eV, respectively. |
Results and Discussion
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WA60089, June 2007
