Lipid-surfactant Mixtures Research Articles

Effect of different polysorbates on development of self-microemulsifying drug delivery systems using medium chain lipids

The primary objective of this study was to develop lipid-based self-microemulsifying drug delivery systems (SMEDDS) without using any organic cosolvents that would spontaneously form microemulsions upon dilution with water. Cosolvents were avoided to prevent possible precipitation of drug upon dilution and other stability issues. Different polysorbates, namely, Tween 20, Tween 40, Tween 60, and Tween 80, were used as surfactants, and Captex 355 EP/NF (glycerol tricaprylate/caprate) or its 1:1 mixture with Capmul MCM NF (glycerol monocaprylocaprate) were used as lipids. Captex 355-Tween-water ternary phase diagrams showed that oil-in-water microemulsions were formed only when the surfactant content was high (80–90%) and the lipid content low (10–20%). Thus, mixtures of Tweens with Captex 355 alone were not suitable to prepare SMEDDS with substantial lipid contents. However, when Captex 355 was replaced with the 1:1 mixture of Captex 355 and Capmul MCM, clear isotropic microemulsion regions in phase diagrams with sizes in the increasing order of Tween 20 < Tween 40 < Tween 60 < Tween 80 were obtained. Tween 80 had the most profound effect among all surfactants as microemulsions were formed with lipid to surfactant ratios as high as 7:3, which may be attributed to the presence of double bond in its side chain that increased the curvature of surfactant layer. Thus, lipid-surfactant mixtures containing 1:1 mixture of medium chain triglyceride (Captex 355) and monoglyceride (Capmul MCM) and as low as 30% Tween 80 were identified as organic cosolvent-free systems for the preparation of SMEDDS. Formulations with a model drug, probucol, dispersed spontaneously and rapidly upon dilution with water to form microemulsions without any drug precipitation.

Assessment of cell viability and permeation enhancement in presence of lipid-based self-emulsifying drug delivery systems using Caco-2 cell model: Polysorbate 80 as the surfactant

PurposeLipid-based self-emulsifying drug delivery systems (SEDDS) are commonly used for solubilizing and enhancing oral bioavailability of poorly water-soluble drugs. However, their effects on viability of intestine epithelial cells and influence on membrane permeation are poorly understood. The present study was undertaken for safety assessment of lipid-based formulations containing medium-chain fatty acid esters as lipids and polysorbate 80 as the surfactant using the Caco-2 in vitro model. Any possible paracellular permeation enhancement through Caco-2 monolayers by the nontoxic formulations was also investigated. MethodsMixtures of monoglyceride (Capmul MCM EP or 708G) or propylene glycol monoester (Capmul PG-8 NF) of medium chain fatty acids with polysorbate 80, with and without the incorporation of a medium-chain triglyceride (Captex 355), were prepared. After suitable dilution with aqueous culture medium, the formulations were incubated with a series of Caco-2 cultures of different maturity. Cell viability and membrane integrity were assessed. Any effects of nontoxic formulations on the transport of the fluorescent dye, Lucifer yellow, through Caco-2 monolayers were also determined. ResultsFormulations containing 1:1 ratios of monoglyceride or propylene glycol monoester to triglyceride (30% polysorbate 80, 35% monoglyceride or monoester and 35% triglyceride) were best tolerated by Caco-2 cells. Increased maturity obtained through longer culture durations rendered Caco-2 cells greater tolerance towards lipid-based formulations, and maximum tolerance to lipid-based formulations was observed with Caco-2 monolayers after being cultured for 21–23days. Furthermore, extent of cell membrane rupture caused by lipid-surfactant mixtures correlated positively with levels of cytotoxicity, suggesting a potential underlying mechanism. Permeation studies using Caco-2 monolayer model revealed that certain formulations significantly enhanced paracellular transport activities. ConclusionsLipid-based SEDDS containing mixtures of monoglyceride (or monoester) and triglyceride of medium chain fatty acids formed fine microemulsions and were significantly less toxic than other formulations. Fully differentiated Caco-2 monolayer was more resistant to lipid-surfactant mixtures than less mature cultures. Certain formulations were also capable of enhancing paracellular permeation.

Cytotoxicity assessment of lipid-based self-emulsifying drug delivery system with Caco-2 cell model: Cremophor EL as the surfactant

PurposeCaco-2 cells are used extensively for in vitro prediction of intestinal drug absorption. However, toxicity of excipients and formulations used can artificially increase drug permeation by damaging cell monolayers, thus providing misleading results. The present study aimed to investigate cytotoxicity of common lipid-based excipients and formulations on Caco-2 cells. MethodsMedium-chain monoglycerides alone or in mixture with the surfactant Cremophor EL, with and without a medium-chain triglyceride, were prepared and incubated with Caco-2 cells from a series of culture stages with varying maturity. Cell viability was evaluated and cell membrane integrity assessed. ResultsCytotoxicity of lipid-based formulations was influenced by the maturity of Caco-2 cells and formulation composition. One-day culture was most sensitive to lipids. When cultured for 5days, viability of Caco-2 cells was significantly improved. The 21-day Caco-2 monolayers maintained the highest survival rate. Microemulsion formulations exhibited significantly less cytotoxicity than neat lipids or surfactant at all stages of cell maturity, and microemulsions containing 1:1 mixtures of monoglyceride and triglyceride appeared to be best tolerated among all the formulations tested. Mechanistically, the observed cytotoxicity was partially due to lipid-induced rupture of cell membrane. ConclusionsMicroemulsions of lipid-surfactant mixtures have less cytotoxicity than lipid alone. Maturity of Caco-2 cells renders significant resistance to cytotoxicity, and monolayers with 21-day maturity are more relevant to in vivo conditions and appear to be a more accurate in vitro model for cytotoxicity assessment.

Simultaneous Formulation, Evaluation and Estimation of Self Emulsifying Drug Delivery System (SEDDS) of Ibuprofen Tablet Dosage form using Varied quantities of Goat Fat as an Excipient and Tween-60 as a Surfactant

The aim of present study confirmed that the SEDDS formulation can be used as a possible alternate to traditional or classical formulations of Ibuprofen to improve its bioavailability using varied quantities of goat fat as an excipient and tween 60 as a surfactant which is a drug in the nonsteroidal anti-inflammatory drug (NSAID) class used for treating pain, fever and inflammation which is also known to be isobutylphenylpropanoic acid. The investigation proved that the formulated dosage forms showed good release profile as well as acceptable tablet properties. The batches with higher tween60: goat fat content ratios gave better release rates. The SEDDS of Ibuprofen was successfully prepared and the tablets prepared passed the various evaluation tests conducted. This method has advantage of reliance on cheap raw materials such as goat fat with fewer processing steps, best suited for lipophilic drugs where the resulting emulsification gives faster dissolution rates and absorption. Invivo evaluation of this novel dosage form is currently in progress. Self-emulsifying drug delivery systems are a promising approach for the formulation of lipophilic drug compounds having poor aqueous solubility. The oral delivery of hydrophobic drugs can be made possible by SEDDSs which have been shown to improve oral bioavailability substantially. From the formulation point of view it is necessary to consider the emulsification properties of lipid base vehicle and the solubility of drug in the lipid surfactant mixture to form a completely miscible solution so as to solubilize adequate quantities of drug in lipid vehicle. Lipids have an important role in absorption process. Thus, SEDDS plays an important role in the formulation of poorly water soluble drugs and enhancing their bioavailability.

Alleviating exercise-induced muscular stress using neat and processed bee pollen: oxidative markers, mitochondrial enzymes, and myostatin expression in rats

BackgroundThe current study was designed to investigate the influence of monofloral Indian mustard bee pollen (MIMBP) and processed monofloral Indian mustard bee pollen (PMIMBP) supplementation on chronic swimming exercise-induced oxidative stress implications in the gastrocnemius muscle of Wistar rats. MethodsMIMBP was processed with an edible lipid-surfactant mixture (Captex 355:Tween 80) to increase the extraction of polyphenols and flavonoid aglycones as analyzed by UV spectroscopy and high performance liquid chromatography-photo diode array. Wistar rats in different groups were fed with MIMBP or PMIMBP supplements at a dose of 100mg/kg, 200mg/kg and 300mg/kg individually, while being subjected to chronic swimming exercise for 4 weeks (5 d/wk). Various biochemical [superoxide dismutase (SOD), glutathione (GSH), malonaldehyde (MDA), nitric oxide (NO), and total protein content], mitochondrial (Complex I, II, III, and IV enzyme activity), and molecular (myostatin mRNA expression) parameters were monitored in the gastrocnemius muscle of each group. ResultsAdministration of both MIMBP (300mg/kg) and PMIMBP (100mg/kg, 200mg/kg, and 300mg/kg) wielded an antioxidant effect by significantly improving SOD, GSH, MDA, NO, and total protein levels. Further MIMBP (300mg/kg) and PMIMBP (200mg/kg and 300mg/kg) significantly improved impaired mitochondrial Complex I, II, III, and IV enzyme activity. Significant down-regulation of myostatin mRNA expression by MIMBP (300mg/kg) and PMIMBP (200mg/kg and 300mg/kg) indicates a muscle protectant role in oxidative stress conditions. ConclusionThe study establishes the antioxidant, mitochondrial upregulatory, and myostatin inhibitory effects of both MIMBP and PMIMBP in exercise-induced oxidative stress conditions, suggesting their usefulness in effective management of exercise-induced muscular stress. Further, processing of MIMBP with an edible lipid-surfactant mixture was found to improve the therapeutic efficiency of pollen.

Complex lipid–surfactant mixtures: a waste of good physical chemistry on a dirty system?

Complex lipid–surfactant mixtures: a waste of good physical chemistry on a dirty system?

Development of Clinical Dosage Forms for a Poorly Water-Soluble Drug II: Formulation and Characterization of a Novel Solid Microemulsion Preconcentrate System for Oral Delivery of a Poorly Water-Soluble Drug

The solution of a poorly water-soluble drug in a liquid lipid–surfactant mixture, which served as a microemulsion preconcentrate, was converted into a solid form by incorporating it in a solid polyethylene glycol (PEG) matrix. The solid microemulsion preconcentrates thus formed consisted of Capmul PG8 (propylene glycol monocaprylate) as oil, Cremophor EL (polyoxyl 35 castor oil) as surfactant, and hydrophilic polymer PEG 3350 as solid matrix. The drug (aqueous solubility: 0.17μg/mL at pH 1–8 and 25°C) was dissolved in a melt of the mixture at 65–70°C and then the hot solution was filled into hard gelatin capsules; the liquid gradually solidified upon cooling below 55°C. The solid system was characterized by differential scanning calorimetry (DSC), scanning electron microscopy (SEM), confocal Raman microscopy (CRM), and the dispersion testing in water. It was confirmed that a solid microemulsion preconcentrate is a two-phase system, where clusters of crystalline PEG 3350 formed the solid structure (m.p. 55–60°C) and the liquid microemulsion preconcentrate dispersed in between PEG 3350 crystals as a separate phase. The drug remained dissolved in the liquid phase. In vitro release testing showed that the preconcentrate dispersed readily in water forming a microemulsion with the drug dissolved in the oil particles (<150nm) and the presence of PEG 3350 did not interfere with the process of self-microemulsification.

Molecular Mechanism of Lateral Diffusion of Fluorosurfactants. A 19F NMR Study

The focus of this thesis has been on the effects of segregation in mixtures of amphiphilic molecules. Two different systems were investigated: fluorocarbon-hydrocarbon surfactant mixtures and lipid-surfactant mixtures.In fluorocarbon-hydrocarbon surfactant mixtures the repulsive interactions between the chains can lead to a demixing into different types of coexisting micelles, fluorocarbon rich and hydrocarbon rich. From NMR self-diffusion measurements such a demixing was found to occur in the mixture of the partially fluorinated surfactant HFDePC and C16TAC. We furthermore suggested a demixing also within the micelles to explain 19F-NMR line width data and results from neutron scattering.In lipid-surfactant mixtures, a segregation of the molecules may instead be caused by a difference in the preferred curvature of the lipid and the surfactant residing within the same aggregate. Using a surfactant selective electrode, binding isoterms of four different cationic surfactants (C12TAC, C14TAC, C16TAC and HFDePC) to preformed lipid (GMO) vesicles were determined. Perforated vesicles were observed by cryo-TEM in the mixture with C16TAC. To explain the results from the binding isoterms, the formation of pores in the bilayer was regarded as a cooperative process, similar to micelle formation. The surfactant accumulates at the edges of the pores, and increasing the surfactant concentration results in an increased number of pores with a constant surfactant/lipid ratio at the edges.The lipid-surfactant mixtures were also studied at the solid/solution interface using AFM. An adsorbed mesh structure, a counterpart to the bulk perforated lamellar phase, was observed for the first time.

Lamellar-to-nematic phase transition in a lipid-surfactant mixture

A lyotropic system, consisting of a lecithin (DMPC) and a non-ionic surfactant (C12E5) in water was studied. The system exhibits a lamellar-to-nematic phase transition. The nematic phase appears as the temperature is decreased and only exists in a very limited temperature and concentration range, for specific lipid-to-surfactant ratios. While a lamellar phase is found at higher temperatures in both mixed and pure C12E5 systems, the transition to the nematic phase at lower temperatures coincides with a micellar phase in the pure C12E5 system. The transition appears to be driven by the strong temperature dependence of the surfactant film spontaneous curvature. The structural properties of the lamellar phase close to the lamellar-to-nematic boundary have been studied by polarised light microscopy and small-angle neutron and X-ray scattering experiments. The signature of a helical defect with Burgers vector of magnitude 2 is apparent in our data, close to the lamellar-to-nematic phase transition. The proliferation of screw dislocations in the lamellar phase might be a plausible mechanism for driving this transition.

NMR Study of Collective Motions and Bending Rigidity in Multilamellar System of Lipid and Surfactant Bilayers

Abstract The frequency dependence of the longitudinal proton spin relaxation time T1 was measured by field-cycling and standard NMR techniques at different temperatures in the liquid crystalline lamellar phases of bilayer systems, composed of lipids, nonionic surfactants, and lipid-surfactant mixtures. We show by our data analysis, comparing various motional models such as layer undulations (LUs) and relaxation by translational diffusion mediated reorientations (TR), that collective layer undulations with their typical T1~ν behaviour determine the low frequency T1 dispersion in both unoriented and glass plate-oriented bilayer systems. The angular dependence of the T1 dispersion for the oriented bilayer system supports these findings and provides a more critical analysis of the two dimensional self-diffusion than in unoriented samples. The evaluated fitting parameters of the LU model allows, together with the measured second moment of the proton NMR signal for the lipid, calculation of the bending rigidity ϰc for these bilayers at different levels of hydration. The obtained values of ϰc turn out to be too large compared with the literature. However, using recent LU models (B. Halle) which include the obvious couplings between neighbouring bilayers at low Larmor frequencies, the corrected ϰc of the fully hydrated membrane systems are comparable to those obtained from the standard videooptical experiments. Therefore proton spin relaxation measure-ments at low Larmor frequencies with the field-cycling technique are a suitable means to determine the bending rigidity ϰc of model membrane systems at low hydrations and of systems containing surfactants.

The heat of transfer of lipid and surfactant from vesicles into micelles in mixtures of phospholipid and surfactant

We study the heat associated with the transformation of vesicles into micelles in mixtures of bilayer-forming phospholipids and micelle-forming surfactants. We subdivide the total heat evolution deltaQ(coex) within the range of coexistence of vesicles and micelles into three contributions related to the transition of dN(D)m-b molecules of surfactant and dN(L)m-b molecules of lipid from micelles to vesicles and to the extraction of dN(D)m-w molecules of surfactant from micelles to the aqueous solution, so that deltaQ(coex) = deltaH(D)m-w x dN(D)m-w + deltaH(D)m-b x dN(D)m-b + deltaH(L)m-b x dN(L)m-b where deltaH(D)m-w, deltaH(L)m-b, and deltaH(D)m-b are the respective molar "transfer" enthalpies. We design a method for the evaluation of all three molar enthalpies, from isothermal calorimetric titrations conducted according to two different protocols of titration of lipid-surfactant mixtures. In the first protocol the mixture is titrated with an aqueous solution of pure lipid vesicles, and in the second the mixture is titrated with an aqueous solution of pure surfactant. Titration of the mixed systems by a buffer solution serves to verify the results obtained under these protocols. In addition to the values of molar enthalpies, our method yields the cmc value of the pure surfactant. We apply our method to investigating the heat evolution in mixtures of egg yolk phosphatidylcholine and the nonionic surfactant octylglucoside in a phosphate-buffered saline solution at 28 degrees C. These studies gave the following values: deltaH(D)m-w = -1732 cal/mol, deltaH(L)m-b = -592 cal/mol, deltaH(D)m-b = 645 cal/mol, and cmc = 23.5 mM. We discuss the possible physical insight of these values and the perspectives of applications of the proposed method.

Comparison of captive and pulsating bubble surfactometers with use of lung surfactants

We compared surface tension measures of surfactants with various surface activities by using a pulsating bubble surfactometer (PBS) and a captive bubble surfactometer (CBS). Rabbit lung lavage surfactant (60,000 x average g for 60 min), bovine surfactant extract (Survanta), and a synthetic lipid surfactant mixture (dipalmitoylphosphatidylcholine-egg phosphatidylglycerol-palmitic acid) were studied at 1.25 mg phospholipid/ml. The PBS was used either unmodified according to manufacturer's instructions or with the sample chamber capillary kept dry and the sample adsorbing at maximum bubble size (5 min). The CBS was used in a manner that imitated the unmodified PBS. We found that all three techniques indicated low surface tension on the first cycle for 60K. For Survanta, the CBS and the modified PBS reported low surface tension on the first cycle, whereas the unmodified PBS did not achieve this within 10 cycles. For the synthetic lipid surfactant mixture, only the CBS measured low surface tension within 10 cycles. Video observations indicate that the modified PBS performs better than the unmodified PBS because keeping the capillary dry prevents surface film from occupying this large surface during cycling, thereby allowing larger area compressions.