Appropiate candidate of pigment source from macroalgae should also contain considerably amount of pigments.
Spectophotometry is fairly accurate method for pigment determination, although shift in absorption peaks occur
frequently. Furthermore, the absorption peaks of different pigments are sometime overlapping each other due to
formation of degradation products such as chlorophyllides and pheophytin which have similar absorption spectra6,33.
These perfect combination may cause ambiguity in pigment determination. Thus, in this study, pigments were also
determined based on literature. Absorption peak around 400 nm to 475 nm were expected as chl a and b. Peak at
around 410 nm and 660 nm might be indicated as pheophytin a while at around 440 nm there was possibility to be
chlorophyllide. Chlorophyta had slight peak at around 470 nm which might be carotenoid. Rhodophyta peaks
indicated phycoerythrin though it was undetected in Gracilaria verrucosa which might be caused by low
concentration. There was considerable peak at around 690 nm that was expected as chl d. Chl d is an exception of
chlorophyll that is dissolve in water and has maximum absorbance at far red region30.
Sample, solvent system, and spectrophotometer might complicate spectrophotometric method25. Cell wall
thickness affects the outcome of maceration. Thicker thallus takes longer time to obtain pigment extract. Gracilaria
verrucosa has fleshy thallus which more difficult to be grinded and produced low concentration of extract. Solvent
selection is very important issue for spectrophotometry since it determines the degree of affinity of solute and
contributes in cell disruption. Combination of certain solvents might give better result of pigment absorbance.
Distilled water can be replaced by phosphate buffer to prevent phycobilin degradation during extracttion24,31. From
this preliminary study, follow up study is highly required in order to get reliable data about macroalgal pigment and
its quantity which use reproducible method.