Table 1 Solid Phase Extraction

Columns

1 ml CN-(?) 100 mg^#

LRC CN-U 100 mg

CN-E 100 mg

LRC CN-E 100 mg

Cond 1

2 ml MeOH

10 ml MeOH

1 ml 0.1% TEA in MeCN

10 ml MeOH

Cond 2

2 ml AmAc^##

5 ml AmAc

2 ml MeOH

2 ml AmAc

Cond 3

None

None

2 ml AmAc

None

Load

1 ml 50% plasma*

Up to 8 ml 50% plasma*

1.1 ml undiluted plasma

2 ml undiluted plasma

Wash 1

2 ml AmAc

10 ml AmAc

2 ml AmAc

2 ml AmAc

Wash 2

2 ml AmAc/20% MeOH

1 ml AmAc/20% MeOH

1 ml water/20% MeOH

1 ml AmAc/20% MeOH

Wash 3

1 ml hexane

5 ml hexane

None

2 ml hexane

Elution

2× 1 ml 0.1% TEA in MeCN

2× 250 μl ethyl acetate

1× 500 μl 0.1% TEA in MeCN

2× 250 μl ethyl acetate

Evap

Nitrogen gas blowdown

Vacuum centrifuge

Nitrogen gas blowdown

Vacuum centrifuge

Injection volume

100 μl (of 200 μl) in MeOH/MeCN/AmAc

90 μl (of 100 μl) in 40% MeCN

50 μl (of 100 μl) in MeOH/MeCN/water

90 μl (of 100 μl) in 40% MeCN