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A novel rapid method for simultaneous determination of three diagnostically important porphyrins in erythrocytes using hyphenated synchronous fluorescence techniques.
[erythropoietic protoporphyria]
The
species
and
concentrations
of
porphyrins
in
erythrocytes
are
of
great
importance
in
the
clinical
screening
and
diagnosis
of
porphyrias
.
However
,
it
is
difficult
to
analyze
them
simultaneously
by
conventional
spectrofluorimetry
.
In
this
paper
,
we
proposed
a
novel
,
simple
and
rapid
method
for
the
simultaneous
determination
of
three
diagnostically
important
porphyrins
in
human
erythrocytes
,
protoporphyrin
IX
(
PP
)
,
coproporphyrin
III
(
CP
)
and
zinc
protoporphyrin
IX
(
ZnPP
)
,
using
hyphenated
techniques
based
on
derivative
matrix
isopotential
synchronous
fluorescence
spectrometry
and
nonlinear
variable-angle
synchronous
fluorescence
spectrometry
(
DMI-NLVASFS
)
.
The
spectral
overlapping
problems
were
well
resolved
and
these
three
components
were
determined
in
one
scanning
without
spectral
compensation
factors
and
chromatographic
separation
.
The
detection
limits
were
0
.
58
,
0
.
21
,
0
.
05
nmol
L
(
-
1
)
for
PP
,
CP
and
ZnPP
,
respectively
.
Only
30
s
was
needed
for
a
single
scanning
and
the
recoveries
were
from
73
%
to
105
%
in
erythrocytes
.
The
Bland-
Altman
analysis
indicates
no
significant
difference
between
the
proposed
DMI-NLVASFS
method
and
conventional
fluorimetry
method
.
The
PP
level
of
the
erythropoietic
protoporphyria
(
EPP
)
patients
was
significantly
higher
than
that
of
healthy
volunteers
.
This
method
can
determine
PP
,
CP
and
ZnPP
simultaneously
in
a
single
scanning
,
thus
providing
a
potential
tool
for
the
clinical
analysis
of
porphyrins
in
human
erythrocytes
and
the
differential
diagnosis
of
porphyrias
.
Diseases
Validation
Diseases presenting
"protoporphyrin ix"
symptom
erythropoietic protoporphyria
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