At 18 days of age, rats from each group were killed
by decapitation, and median eminences (ME) and
spleens were rapidly removed and cooled on ice. The
tissues were treated with perchloric acid (0.1 M) and
disrupted by homogenization. Alumina was added to
the tissue to absorb the catecholamines, and the samples
were washed and eluted in perchloric acid (0.1 M)
using a microfilter kit. Aliquots of the eluted extract
were subjected to high-performance liquid chromatography
(HPLC) with electrochemical detection to determine
NA and metabolite levels. The concentration of
the metabolite 3-methoxy-4-hydroxyphenylglycol
(HMPG) was measured in ME only. For chromatographic
separations, a Biophase ODS (5 pm) analytical
column was used. The mobile phase was a mixture of
0.15 M monochloroacetate, pH 3.0 with 2.0 mM
Na,EDTA and 28 mg/l sodium octyl sulfate. AI1 solutions
were filtered and degassed. The loo-p1 injection
was made through a fixed loop (20 ~1) injection valve.
Flow rates of 1.3-1.5 ml/min
At 18 days of age, rats from each group were killed
by decapitation, and median eminences (ME) and
spleens were rapidly removed and cooled on ice. The
tissues were treated with perchloric acid (0.1 M) and
disrupted by homogenization. Alumina was added to
the tissue to absorb the catecholamines, and the samples
were washed and eluted in perchloric acid (0.1 M)
using a microfilter kit. Aliquots of the eluted extract
were subjected to high-performance liquid chromatography
(HPLC) with electrochemical detection to determine
NA and metabolite levels. The concentration of
the metabolite 3-methoxy-4-hydroxyphenylglycol
(HMPG) was measured in ME only. For chromatographic
separations, a Biophase ODS (5 pm) analytical
column was used. The mobile phase was a mixture of
0.15 M monochloroacetate, pH 3.0 with 2.0 mM
Na,EDTA and 28 mg/l sodium octyl sulfate. AI1 solutions
were filtered and degassed. The loo-p1 injection
was made through a fixed loop (20 ~1) injection valve.
Flow rates of 1.3-1.5 ml/min
การแปล กรุณารอสักครู่..