Biomedical Engineering Reference
In-Depth Information
Organic solvent
a
volume ratio
b
Steps prior to LLE Recovery (%) Reference
Et
2
O/H
2
CCl
2
2:1 Hum plasma Trop 1:3.4 +IS +PB, pH 10.0 72-77 [
83
]
Et
2
O/H
2
CCl
2
7:3 Hum plasma Gran 1:9.5 +IS 63 [
73
]
Me-O-
t
Bu Hum plasma Scp n.s. +IS +alkalized n.s. [
88
]
For abbreviation of analyte names see Sect. “Abbreviations”.
ACN
acetonitrile,
b -glucu
solution of b -glucuronidase (
E. coli
),
EE
ethyl acetate,
hep-SA
heptane-
sulfonic acid,
iso
prop
iso-propanol,
IS
internal standard,
MeOH
methanol,
metab.
metabolites,
Me-O-
t
Bu
, methyl-
t
butyl ether;
n.s.
not speci fi ed,
OAc
acetate,
PB
phosphate buffer
a
Numbers give volume ratio v/v
b
Numbers in parentheses give the number of repeated LLE cycles
c
A second LLE step followed by adding 0.1 M PB, pH 7.4 to the organic layer (ratio n.s.)
d
Microsomes from rat, mouse, hamster, guinea pig
e
A second LLE step followed by adding 0.5 M HCl to the organic layer (1:4)
f
After the first LLE step a number of additional extraction steps followed: the organic layer of first LLE was mixed with 0.1 M HCl (1:0.38) for second LLE.
EE was added to that acidic aqueous layer (1:3) for third LLE. That aqueous layer was alkalized (pH 9.0) with 25 % NH
4
OH and mixed with H
2
CCL
2
(1:2.7)
for fourth LLE. That organic phase was evaporated to dryness, re-dissolved and analysed by LC-APCI MS/MS
g
After the first LLE step an additional step followed: potassium iodide, glycine, NaOH, and NaCl were added to the aqueous layer prior to second and third
LLE with H
2
CCl
2
(1:0.7). The latter organic layers were evaporated to dryness and analysed by LC-ESI MS.
Sample matrix
Analyte
