2.4. Analysis of taurine content
Egg and larvae samples were rinsed in distilled water and then
lyophilized (Freeze dryer FD-1C-80, Boyikang, China). About 2 to 5 mg
from each sample were first hydrolyzed (1 ml of 0.1 M HCL and homogenized
for 1.5 min on ice) and then centrifuged (13,000 rpm at 4 °C for
20 min). A measured amount of the supernatant from each sample was
transferred into 10 ml glass test tubes. Further preparation and analysis
of samples were conducted according to the procedures described by
Orth (2001), using reversed-phase HPLC (Varian HPLC 325–410 nm,
Walnut Creek, California, USA) with gradient elution separation on a
150 × 4.6 mm Dionex C18 5 μm column. An adjustment in the gradient
elution program was made in order to optimize separation and better
detection of taurine in the sample.