Drug Metabolism in Drug Design and Development Basic Concepts and Practice

TABLE 16.1 LC–MS/MS analyses of individual CYP marker substrates in pooled human liver microsomes.CYPinvolved

Substrate

LC column

LC gradient

MetaboliteQ

1 /Q

3 mass (

m

/z

)

Internal standardQ

1 /Q

3 mass (

m

/z

)

CYP1A2

Phenacetin

AquaSep, 5

m

m,

2.0

50 mm

5–50% B

1.4 min linear

Acetaminophen

151.9/110.1

(^04)
-OH-Butyranilide180.2/71.0
CYP2A6
Coumarin
Zorbax SBC8,
3.5
m
m,
4.6
75 mm
15–65%
7 min linear
7-OH-Coumarin
HPLC
CYP2B6
Bupropion
BDS Hypersil
C8, 5
m
m,
2
50 mm
20–80% B
1.9 min linear
OH-Bupropion
256.2/184.2
dl-Propranolol
260.2/155.1
Diazepam
Extend-C18,
3.5
m
m,
4.6
50 mm
30–70% B
7.0 min linear
Nordiazepam
300.9/255.2
Lorazepam
320.9/275.2
CYP2C8
Taxol
Zorbax 300 Extend
C18, 5
m
m,
4.6
50 mm
20–80% B
2.0 min linear
6 a
-OH Taxol870.3/525.2
Baccatin III
587.2/405.2
CYP2C9
Diclofenac
BDS Hypersil
C8, 5
m
m,
2
50 mm
10–70% B
1.9 min linear
(^04)
-OH-Diclofenac312/231.1
Flufenamic acid
282.1/264.1
CYP2C19
(S
)-Mephenytoin
Zorbax SB-Aq,
5
m
m, 4.6
50 mm
20–80% B
1.9 min linear
(^04)
-OH-Mephenytoin235.1/150.1
Phenytoin
253.2/182.2
CYP2D6
Dextromethorphan
XDB C8, 5
m
m,
2.1
50 mm
Agilent
0–60% B
2.1 min linear
Dextrorphan
258.2/157.0
Levallorphan
284.1/199.2
518