In the present study, a simple and rapid HPLC procedure has been developed for determination of a wide range of lactic acid concentrations in blood serum from edible crab (Cancer pagurus). The lactic acid concentration is used as one of several stress indicators in edible crab.
During the lactic acid analysis, the analytic column, Ace 5 C18, was operating at 20ºC. The mobile phase, 10 mM KH2PO4:acetonitrile (99:1), pH 2.5, was delivered isocratic, at a flow rate of 0.2 ml/min, in a 10 min run. Detection and identification were performed using a photodiode array detector at 220 nm. Detection limits were found in the range of 0.02 mM (s/n=8) per 10 μl sample injected, while linearity was measured up to 118 mM. Statistical evaluation of the method was examined performing intra-day (n=10) (VC% = 0.3-3.3%) and inter-day (n=5) (VC% = 0.0-1.3%) calibration and was found to give high accuracy and precision results.
Blood serum samples previously analysed by Vitros 950 chemistry station, were analysed by the developed HPLC method (r=0.94).
