In fact, it is possible to confirm that between K2HPO4 and K3PO4, the latter inorganic salt has the highest capacity to induce the phase separation, although in some cases, only a small difference is observed. This behaviour can be easily supported by literature data and it is related to the idea that the strong salting-out inducing anions, PO43− and HPO42−, exhibit a stronger capability for creating ion-hydration complexes by excluding

water from the alcohol-rich phase, and thus favouring the formation of ATPS (He, Li, Liu, Li, & Liu, 2005). Also, according to literature, the K2HPO4/KH2PO4 salts have a lower ability for the ATPS formation, due to the presence of KH2PO4, which tends towards the salting-in regime. Indeed, Metformin it was already described that KH2PO4 is not capable by itself to promote the formation of alcohol-based ATPS. Here the “usual” behaviour of K2HPO4/KH2PO4 was only detected

for the 1-propanol system. Searching for an PI3K inhibitor review explanation for this behaviour, the pH of both phases of each system were measured (Table 1). According to Table 1, it is observed that the pH is salt-dependent and alcohol-independent. The addition of some of these alcohols is responsible for the destruction of the buffer condition, which is demonstrated by significant differences in the expected pH values of the phases. The buffer condition was lost in most of the systems, with the exception of the 1-propanol. Thus, for the ternary systems with K2HPO4/KH2PO4 and methanol, ethanol

and 2-propanol, the effect is not driven by the phosphate buffer ionic strength and respective interactions, but it is induced by the presence of two different inorganic salts, K2HPO4 and KH2PO4, as individual ionic species, and which partition in different directions of the system. Since ATPS making use of K2HPO4/KH2PO4 were not found in literature, a comparison between our results and those in the literature was not possible. Evidently, the use of these ternary systems for extraction oxyclozanide purposes should be cautiously carried out since the pH value is not neutral for systems composed of methanol, ethanol or 2-propanol. The solubility curves described before, were correlated using the mathematical approach originally described in literature (Merchuck et al., 1998), by the application of Eq. (1). The regression parameters A, B and C, the respective standard deviations (std), and the correlation coefficients (R2), are reported in Table S6 in Supporting Information. To complete the phase diagrams, the tie-lines (TLs), and respective tie-line lengths (TLLs), were determined. Their values are reported in Table S7 in Supporting Information, along with the compositions of inorganic salt and alcohol at the top (T) and bottom (B) phases. The graphical representation of the phase diagrams of all the systems studied is depicted in Supporting Information ( Figures S2 to S12).