Niflumic Acid/Camphorsulfonic Acid Extracted from Rat Plasma

Plate:

Oasis WAX μElution Plate

Condition:

200 μL MeOH

Equilibrate:

200 μL H₂O

Load:

100 μL 1:1 diluted rat plasma, 10 pg/μL, w/2% H₃PO₄

Wash 1:

200 μL 25 mM NH₄CH₂COOH buffer in H₂O, pH 5

Wash 2:

200 μL MeOH

Elute:

50 μL (25 μL x 2) MeOH w/2% NH₄OH

Dilute:

50 μL H₂O with 2% HCOOH

Inject:

10 μL

Column:

SunFire C₁₈ 2.1 x 20 mm IS, 3.5 μm

Part number:

186002531

Mobile phase A:

H₂O with 10 mM NH₄CH₃COOH, pH 5.5

Mobile phase B:

MeOH with 10 mM NH₄CH₃COOH, pH 5.5

Flow rate:

0.4 mL/min

Injection volume:

10 μL

Sample concentration:

10 pg/μL each analyte

Temperature:

Ambient

Instrument:

Waters Alliance 2695

Waters Micromass Quattro micro API

ESI source temp.:

150 °C

Desolvation temp.:

350 °C

Cone gas flow:

50 L/Hr

Desolvation gas flow:

600 L/Hr

Collision Cell:

1.23e^-3 bar (Ar gas)

Compounds

MRM Transition

Cone (V)

CID(eV)

Niflumic Acid

m/z 281.1 → 237.1

30

17

Camphorsulfonic Acid

m/z 231.1 → 79.9

45

30