Optimized LC–MS/MS quantification method for the detection of piperacillin and application to the development of charged liposaccharides as oral penetration enhancers

In vitro and In vivo evaluation of positively charged liposaccharide derivatives as oral absorption enhancers for the delivery of anionic drugs

“…5) were calculated from concentrations of piperacillin determined by liquid chromatography coupled to mass spectrometry (LC-MS/MS) against an eight point standard curve while 14 C-mannitol was quantified by liquid scintillation counting (b-emission). 10 Piperacillin alone expectedly showed very low permeability across Caco-2 monolayers; the observed values were higher than reported previously, 3.07(OH)-3.22(Na) Â 10 À7 cm s À1 versus 3.24(Na) Â 10 À8 cm s À1 . 10 A similar 10-fold variation was noticed for Pip-GC 12 [1:1] and 14 C-mannitol, this variability could be explained by the difference in cell passage numbers, since the cells used in the present study were much younger than in the previous examination.…”

“…10 Surprisingly, the penetration of piperacillin was not significantly improved when combined with GC 12 in a 1:2 molar ratio. No satisfactory ITC data could be obtained for GC 12 so the ratio for optimal binding could not be assessed accurately.…”

“…For LC-MS/MS experiments, a Phenomenex luna C18 column (5 mm, 50 mm  2.0 mm) equipped with a C18 guard column (4 mm  3.0 mm) was attached to the mass spectrometer and a 30-100% gradient of B in A over 4 min was used for elution at a flow rate of 0.5 mL/min with a 1:10 splitter upstream from the ionisation source (Shimadzu LC-10AT system). 10 Data were acquired with Analyst 1.4 software (Applied Biosystems/MDS Sciex, Toronto, Canada).…”

“…8 Our recent investigations have demonstrated the potential of charged glycolipids to form ion-pairs with a model drug like piperacillin. 9,10 In an earlier study, we reported that the in vitro permeability of piperacillin across Caco-2 monolayers could be notably improved by associating this anionic b-lactam antibiotic to a positively charged liposaccharide containing a D-glucose moiety coupled to a 12-carbon long lipoamino acid (LAA) (a-amino acid with lipophilic alkyl side-chain). 10 Positive results were also observed when varying the lipid chain length.…”

Design and synthesis of a series of novel, cationic liposaccharide derivatives as potential penetration enhancers for oral drug delivery

“…8 Significant increases in apparent permeability coefficients were observed, notably with a glucose-based derivative containing a 12-carbon long lipid chain, however the permeability values were still low (<10 À6 cm s À1 ), indicating that if administered orally, these formulations would unlikely be fully absorbed. 9 Furthermore, the pH gradient residing in the gastro-intestinal tract might cause the drug-glucolipid entities to dissociate as one of the residue could be partially neutralized, negatively affecting the permeability enhancing effect of the designed glucolipids.…”