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Performance enhancement in the measurement of 5 endogenous steroids by LC-MS/MS combined with differential ion mobility spectrometry.
[congenital adrenal hyperplasia]
Challenges
for
steroid
analysis
by
LC
-
MS
/
MS
include
low
ionization
efficiency
,
endogenous
isobars
with
similar
fragmentation
patterns
and
chromatographic
retention
.
Differential
ion
mobility
spectrometry
(
DMS
)
provides
an
additional
degree
of
separation
prior
to
MS
/
MS
detection
,
and
shows
promise
in
improving
specificity
of
analysis
.
We
developed
a
sensitive
and
specific
method
for
measurement
of
corticosterone
,
11
-
deoxycortisol
,
11
-
deoxycorticosterone
,
17
-
hydroxyprogesterone
and
progesterone
in
human
serum
and
plasma
using
an
ABSciex
5500
mass
spectrometer
equipped
with
a
differential
ion
mobility
interface
.
250
μL
aliquots
of
serum
were
spiked
with
deuterated
internal
standards
and
extracted
with
MTBE
.
The
samples
were
analyzed
using
positive
mode
electrospray
LC
-DMS-
MS
/
MS
.
The
method
was
validated
and
compared
with
immunoassays
and
LC
-
MS
/
MS
methods
of
reference
laboratories
.
Inter
and
intra
assay
imprecision
was
<
10
%
.
Limits
of
quantification
and
detection
in
nmol
/
L
were
0
.
18
,
0
.
09
for
corticosterone
and
17
-
hydroxyprogesterone
,
0
.
30
,
0
.
16
for
11
-
deoxycortisol
,
0
.
12
,
0
.
06
for
progesterone
and
0
.
06
,
0
.
03
for
11
-
deoxycorticosterone
.
Comparison
for
progesterone
and
17
-
hydroxyprogesterone
with
immunoassay
showed
slopes
of
0
.
97
and
1
.
0
,
intercepts
of
0
.
16
and
0
.
10
and
coefficients
of
determination
(
r
(
2
)
)
of
0
.
92
and
0
.
97
,
respectively
.
Progesterone
by
immunoassay
showed
positive
bias
in
samples
measuring
<
3
.
18
nmol
/
L
.
Reference
intervals
for
progesterone
and
11
-
deoxycorticosterone
in
post-menopausal
women
were
found
to
be
<
2
.
88
and
<
0
.
28
nmol
/
L
respectively
.
We
developed
and
validated
an
LC
-DMS-
MS
/
MS
method
for
analysis
of
five
endogenous
steroids
suitable
for
routine
measurements
in
clinical
diagnostic
laboratories
.
Specificity
gained
with
DMS
allows
reducing
the
complexity
of
sample
preparation
,
decreasing
LC
run
times
and
increasing
speed
of
the
analysis
.
Diseases
Validation
Diseases presenting
"endogenous isobars with similar fragmentation patterns and chromatographic retention"
symptom
congenital adrenal hyperplasia
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