Biomedical Engineering Reference
In-Depth Information
Table 6
(continued)
An LC/ESI
+
-MS/MS
method for estrogens in
plasma following
IUPAC guideline [88]
FDA Guidance for bioanalytical
method validation (http://fda.
gov/edcr/guidance/index.htm)
EPA Method 539—7 hormones in
drinking water by LC/ESI
+/−
-MS/
MS (http://water.epa.gov/drink/)
LC/ESI
+
-MS/MS methods for
hormones in serum following NIST
297 and ISO15193 guidelines [84, 85]
Parameter
Sample
stability
3× Freeze-thaw cycles for
samples between −70 °C
and room temperature;
4-24 h at room temperature;
long-term storage; sample
stock and postpreparation
stability.
5-10 ng/L × 6 at £ 6 °C for 28 days
NA
3× Freeze-thaw cycles
for samples between
−20 °C and room
temperature;
Ambient and 4 °C for
7 days
Internal
standard
Isotope labeled analyte with
optimized binding
assessment, and verified
standard curve
IS peak area within ±50 % of each
analyte
13
C
6
-Estradiol, 16 a -Hydroxyestra-
diol-
d
2
, Testosterone-
d
3
,
13
C
2
-Etheylnylestradiol
IS peak area within ± 50 % of each
analyte
Testosterone-
d
3
, Estradiol-
d
3
Estron-d
4
, Estradiol-d
5
Sample
prepara-
tion
Extraction recovery (at three
levels) of analyte and
internal standard should be
consistent, precise, and
reproducible.
SPE, without derivatization
SPE + LLE (³80 %), no derivatization
for testosterone, Estrogens
derivatized with dansyl chloride
LLE (100 %), deriva-
tized with dansyl
chloride
