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A novel HPLC-electrochemical detection approach for the determination of D-penicillamine in skin specimens.
[cystinuria]
D-
penicillamine
is
a
thiol
drug
mainly
used
for
Wilson
's
disease
,
rheumatoid
arthritis
and
cystinuria
.
Adverse
effects
during
normal
use
of
the
drug
are
frequent
and
may
include
skin
lesions
.
To
evaluate
its
toxic
effects
in
clinical
cases
an
original
method
based
on
high
performance
liquid
chromatography
coupled
to
amperometric
detection
in
a
specific
biological
matrix
such
as
skin
has
been
developed
.
The
chromatographic
analysis
of
D-
penicillamine
was
carried
out
on
a
C
18
column
using
a
mixture
of
acid
phosphate
buffer
and
methanol
as
the
mobile
phase
.
Satisfactory
sensitivity
was
obtained
by
oxidizing
the
molecule
at
+
0
.
95
V
with
respect
to
an
Ag
/
AgCl
reference
electrode
.
A
chemical
reduction
of
D-
penicillamine-protein
disulphide
bonds
using
dithioerythritol
combined
with
microwaves
was
necessary
for
the
determination
of
the
total
amount
of
D-
penicillamine
in
skin
specimens
.
A
further
solid-phase
extraction
procedure
on
C
18
cartridges
was
implemented
for
the
sample
clean-up
.
The
whole
analytical
procedure
was
validated
:
high
extraction
yield
(
>
91
.
0
%
)
and
satisfactory
precision
(
RSD
<
6
.
8
%
)
values
were
obtained
.
It
was
successfully
applied
to
skin
samples
from
a
patient
who
was
previously
under
a
long
-term
,
high
-dose
treatment
with
the
drug
and
presented
serious
D-
penicillamine-related
dermatoses
.
Thus
,
the
method
seems
to
be
suitable
for
the
analysis
of
D-
penicillamine
in
skin
tissues
.
Diseases
Validation
Diseases presenting
"skin samples from a patient"
symptom
cystinuria
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