Application of capillaries with minimized electroosmotic flow to the electrokinetic study of acidic drug–β-oleoyl-γ-palmitoyl-l-α-phosphatidyl choline liposome interactions

Interaction of a model set of common drugs varying widely in their polarity as well as in their chemical structure (salicylic acid, acetylsalicylic acid, ketoprofen, phenytoin and propranolol) with β-oleoyl-γ-palmitoyl-l-α-phosphatidyl choline (POPC) liposomes was investigated by means of capillary electrophoresis. Two phosphate buffers differing in their pH (50 mM, pH 7.5 and 9.2) were used both for liposome reconstitution and as background electrolytes for capillary electrophoresis using capillaries with minimised electroosmotic flow (EOF). The liposomes showed practically no electrophoretic mobility and formed a stable plug in the capillary. At alkaline pH (9.2), the polyimide coated capillary exhibited residual endoosmotic flow (the EOF marker appeared before the detection window around 40 min as compared to 2.2 min in the untreated capillary; attempts to reveal endoosmotic flow at pH 7.5 were unsuccessful). The concentration of the mixture of the test compounds was 50 μg/ml (except for ketoprofen concentration of which was 5 μg/ml due to the lower solubility of the drug), i.e. large enough to exceed the binding capacity of the injected liposome plug at least at the neutral pH (7.5) which consequently resulted in two regions in the electropherogram, namely that which contained the unbound species and that corresponding to the liposome (lipid)-bound fraction. On the other hand in runs done at high pH of the background electrolyte (9.2) the whole amount injected interacted with the liposomes. Acidic drugs and phenytoin were run with negative polarity at the injection site. It was documented that both at pH 7.5 and 9.2 the investigated solutes interacted with POPC liposomes, though at pH 7.5 the equilibrium between the bound and unbound drugs was in favor of the unbound species. On the contrary, at pH 9.2 binding was considerably stronger and only the liposome bound fraction was seen upon electrophoresis. The well-known instability of phenytoin at room temperature resulted in the formation of an acidic hydrolytic product which was strongly bound to liposomes at the higher pH value. While no binding of phenytoin could be established at pH 7.5, at pH 9.2 this compound was degraded (hydrolyzed) and its degradation product was clearly bound to liposomes. It has to be emphasized that binding experiments must be done separately for acidic/neutral and basic drugs; binding of acidic/neutral drugs must be done at reversed polarity, while in order to reveal binding of basic drugs, positive polarity at the injection site must be used.