For measurement of hemocyanin,100microlitre hemolymph were immediately diluted with 900 microlitre distilled water in a 10-mm quartz cuvette and the absorbance was measured at 335nm (characteristic of oxyhemocyanin) usinga Hitachi U-2000 spectrophotometer (Hitachi Ltd., Tokyo, Japan). Using an extinction coefficient of 17.26, on the basis of functional subunit of 74000(Antonini and Brunori, 1974),the hemocyanin concentration was calculated.

{Cheng & Chen}

But, recent papers suggests that take 10 micro litre Haemolymph and 990 micro litre DDW and read at 335nm, Using an extinction coefficient of 17.26, on the basis of functional subunit of 74000(Antonini and Brunori, 1974),the hemocyanin concentration was calculated.