Comparison of in vitro dialysis release methods of loperamide-encapsulated liposomal gel for topical drug delivery

Abstract

Background: The purpose of this study was to determine the most appropriate dialysis equilibrium method to assess liposomal gel formulations containing hydrophobic drugs, to give the most accurate indication of drug release.

Methods: Loperamide hydrochloride-encapsulated liposomes, composed of L-α-phosphatidylcholine and cholesterol (molar ratio of 2:1), were prepared according to the method of dried lipid film hydration. The liposomes were incorporated into a carbopol gel (0.5%, weight/weight). The release of the drug from the nanoparticles was assessed using a number of variations of the dialysis technique, taking into account solubility parameters and formulation. Method 1 (below saturation point) and Method 2 (above saturation point) used a dilution method to evaluate how drug concentration and solubility affects the in vitro drug-release profile of loperamide hydrochloride, while Methods 3 (below saturation point) and 4 (above saturation point) evaluated how drug concentration and the gel base affect the release profile.

Results: In Method 1, the liposomes showed a rapid release of just over 60% in the first 3 hours and then a slower, sustained release to just over 70% at 24 hours. Method 2 showed a gradual, sustained release profile with the liposomes with 55% release at 24 hours. In Method 3, the liposomes showed a rapid burst release of 98% at 2 hours. In Method 4, the liposomal gel had a rapid release of 60% within 3 hours and then a more gradual, sustained release with 86% release at 24 hours. The free drug suspension in Methods 2 and 4 showed a limited release across the dialysis membrane, in comparison to Methods 1 and 3, which showed a complete release in a timely manner.

Conclusion: This study has demonstrated that the actual method used for equilibrium dialysis plays a significant role in determining the true characteristics of a topical nanoformulation, with Method 3 providing the most accurate indication of the release of a hydrophobic drug from a topical liposomal formulation.

Keywords: gel; in vitro drug release; liposomes; loperamide; topical; transdermal.

Figure 1

Method 1 (1:4 dilution). Notes: In vitro release of loperamide HCl in PBS (pH 6.5) for liposomal and free drug solution. Values are expressed as mean ± standard deviation; n=3 independent experiments. Abbreviations: HCl, hydrochloride; PBS, phosphate buffered saline.

Figure 2

Method 1 (1:10 dilution). Notes: In vitro release of loperamide HCl in PBS (pH 6.5) for liposomal and free drug solution. Values are expressed as mean ± standard deviation; n=3 independent experiments. Abbreviations: HCl, hydrochloride; PBS, phosphate buffered saline.

Figure 3

Method 2 (1:4 dilution). Notes: In vitro release of loperamide HCl in PBS (pH 6.5) for liposomal and free drug suspension. Values are expressed as mean ± standard deviation; n=3 independent experiments. Abbreviations: HCl, hydrochloride; PBS, phosphate buffered saline.

Figure 4

Method 2 (1:10 dilution). Notes: In vitro release of loperamide HCl in PBS (pH 6.5) for liposomal and free drug suspension. Values are expressed as mean ± standard deviation; n=3 independent experiments. Abbreviations: HCl, hydrochloride; PBS, phosphate buffered saline.

Figure 5

Method 3. Notes: In vitro release of loperamide HCl in PBS (pH 6.5) for liposomal gel and free drug solution in gel. Values are expressed as mean ± standard deviation; n=3 independent experiments. Abbreviations: HCl, hydrochloride; PBS, phosphate buffered saline.

Figure 6

Method 4. Notes: In vitro release of loperamide HCl in PBS (pH 6.5) for liposomal gel and free drug suspension in gel. Values are expressed as mean ± standard deviation; n=3 independent experiments. Abbreviations: HCl, hydrochloride; PBS, phosphate buffered saline.