Construction of Phase Diagram

The phase diagram is constructed using cloud-point method at room temperature and atmospheric
pressure. The aqueous solution of IL (from 60 wt% to 80 wt%) and salts solution (≈40 wt%) are
prepared gravimetrically (± 10−4 g) and are used for determination of the saturation curves. The
salts solution is added into each IL solution dropwise until observation of cloudy solution in which
two phases are formed, followed by addition of distilled water until detection of the monophasic
regime. The experiment is conducted under constant stirring. Each mixture composition is
identified by measuring the weight of the components added using an analytical balance. The final
composition of IL and salts are calculated and binodal curves of IL-ATPS are plotted by varying
concentrations of IL (Chang et al., 2018 ;Quental et al., 2015).

The tie lines are determined by gravimetric method as proposed by Merchuck et al. Experimental
binodial curve are correlated by least-square regression using non-linear eq1:
IL = A exp [B (Salt)0.5 – C (Salt)3]
Where IL and salts are the mass fraction percentages and A,B, C are constants obtained by data
regression
Crude Extraction
Dried S. platensis powder is suspended in distilled water (phosphate buffer 0.1M = pH7) at the solid
to volume ratio of 1:15 (w:v) (Luo et al., 2016).

The mixture is then homogenized using Ultraturrax (IKA dispersion S18N-19G) at 13500rpm for 15min
to disrupt the cell wall of microalgae thus releasing the phycobiliprotein (Santos et al., 2018) in an ice
bath. Next, the mixture is centrifuged at 10000rpm for 15min at 4C (Izadi and Fazilati, 2018). The
resulting supernatant is separated carefully using syringe. The concentration of CPC, total protein and
carbohydrate are analysed and the supernatant is stored at 4ºC for further use.