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A column-switching HPLC-MS/MS method for mucopolysaccharidosis type I analysis in a multiplex assay for the simultaneous newborn screening of six lysosomal storage disorders.
[krabbe disease]
Lysosomal
storage
disorders
comprise
a
group
of
rare
genetic
diseases
in
which
a
deficit
of
specific
hydrolases
leads
to
the
storage
of
undegraded
substrates
in
lysosomes
.
Impaired
enzyme
activities
can
be
assessed
by
MS
/
MS
quantification
of
the
reaction
products
obtained
after
incubation
with
specific
substrates
.
In
this
study
,
a
column-switching
HPLC-
MS
/
MS
method
for
multiplex
screening
in
dried
blood
spot
of
the
lysosomal
enzymes
activities
was
developed
.
Mucopolysaccharidosis
type
I
,
Fabry
,
Gaucher
,
Krabbe
,
Niemann-
Pick
A
/
B
and
Pompe
diseases
were
simultaneously
assayed
.
Dried
blood
spots
were
incubated
with
substrates
and
internal
standards
;
thereafter
,
supernatants
were
collected
with
minor
manipulations
.
Samples
were
injected
,
trapped
into
an
online
perfusion
column
and
,
by
a
six
-port
valve
,
switched
online
through
the
C
18
analytical
column
to
perform
separation
of
metabolites
followed
by
MS
/
MS
analysis
.
A
total
of
1136
de
-identified
newborn
screening
samples
were
analyzed
to
determine
references
for
enzymes
activity
values
.
As
positive
controls
,
we
analyzed
dried
blood
spots
from
three
patients
with
Pompe
,
one
with
Fabry
,
one
with
Krabbe
disease
and
two
with
MPS
I
,
and
in
all
cases
the
enzyme
activities
were
below
the
cutoff
values
measured
for
newborns
,
except
for
an
MPS
I
patient
after
successful
hematopoietic
stem
cell
transplantation
.
Diseases
Validation
Diseases presenting
"rare genetic diseases"
symptom
classical phenylketonuria
fabry disease
krabbe disease
primary hyperoxaluria type 1
severe combined immunodeficiency
werner syndrome
wiskott-aldrich syndrome
x-linked adrenoleukodystrophy
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