This is an Application Brief and does not contain a detailed Experimental section.
This application brief demonstrates the analysis of cephalexin on Oasis MCX.
Cephalexin is a member of the cephalosporin class of antibiotics used for the treatment of bronchitis, pneumonia, UTI’s and infections of the ears and throat. This molecule is zwitterionic, meaning it has both carboxylic acid and amine moieties. Because the molecule has ionizable functionalities that can be either positively or negatively charged, screening all 4 Oasis sorbents following the Oasis 2x4 Method is clearly the mechanism for identifying the best sorbent. Oasis MCX resulted in the best recoveries and would be used for further sample preparation method development.
Oasis MCX 10-mg plate ( P/N 186000259) |
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Condition: |
500 μL MeOH |
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Equilibrate: |
500 μL H2O |
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Load: |
500 μL (250 μL rat plasma, diluted 1:1 with 4% H3PO4 in H2O) |
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Wash 1: |
500 μL 2% HCOOH |
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Wash 2: |
500 μL MeOH |
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Elute: |
250 μL (125 μL x 2) 5% NH4OH in MeOH |
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Options: |
1. Dilute 250 μL H2O with 2% FA 2. Evaporate/ Reconstitute 3. Direct inject |
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Inject: |
10 μL |
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Column: |
ACQUITY UPLC BEH C18 2.1 x 50 mm, 1.7 μm |
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Mobile phase A: |
0.1% HCOOH in H2O |
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Mobile phase B: |
0.1% HCOOH in MeOH |
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Flow rate: |
0.4 mL /min |
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Injection volume: |
10.0 μL |
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Column temperature: |
45 °C |
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Sample temperature: |
15 °C |
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Sample diluent: |
50/50 water/methanol |
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Instrument: |
ACQUITY UPLC with Quattro Premier |
ESI+ capillary: |
3.0kV |
Source temp.: |
120 °C |
Desolvation temp.: |
350 °C |
Cone gas flow: |
50 L /Hr |
Desolvation gas flow: |
700 L /Hr |
Collision cell pressure: |
2.59 e-3 mbar |
MRM transition: |
348 → 157.9 |
Cone voltage: |
20V |
Collision energy: |
18eV |
WA60085, June 2007