Diffusion cell absorption

The two media typically used include Simulated Gastric Fluid (pH 1-pH 3) and Simulated Intestinal Fluid (pH 6-pH 7). The drug substance under investigation is introduced, and its uptake in the diffusion cell ( absorption ) is governed by its hydrophilic-lipophilic balance (HLB). The absorption model proposed by Strieker (26) in the early 1970s therefore effectively took into consideration (in an experimental sense) all aspects considered by the theory of the BCS, which was introduced more than 20 years later. 

The Sartorius Absorption Model (26), which served as the forerunner to the BCS, simulates concomitant release from the dosage form in the GI tract and absorption of the drug through the lipid barrier. The most important features of Sartorius Absorption Model are the two reservoirs for holding different media at 37°C, a diffusion cell with an artificial lipid barrier of known surface area, and a connecting peristaltic pump which aids the transport of the solution or the media from the reservoir to the compartment of the diffusion cell. The set-up is shown in Figures 7a and b. 

In many cases, transdermal drug absorption is investigated using a Franz-diffusion cell. The concentrations both in the membrane and the acceptor compartment are assumed to be zero at the start of the experiment. At different time points, the cumulative drug amount per unit area in the receptor q(t) is determined and plotted versus time t (Figure 20.1). After some time, the flux... 

OECD has adopted an in vitro test for skin absorption potential (OECD TG 428, Skin Absorption In Vitro Method). According to this guideline, excised skin from human or animal sources can be used. The skin is positioned in a diffusion cell consisting of a donor chamber and a receptor chamber, between the two chambers. The test substance, which may be radio-labeled, is applied to the surface of the skin sample. The chemical remains on the skin for a specified time under specified conditions, before removal by an appropriate cleansing procedure. The fluid in the receptor chamber is sampled at time points throughout the experiment and analyzed for the test chemical and/or metabolites. 

Absorption of phenol in a flow-through diffusion cell in vitro, using full-thickness... 

Van Zyl et al. reported on the diffusion of ipratropium through porcine bronchial epithelium tissue [74], In principle, ipratropium is administered via the respiratory tract by inhalation to treat pulmonary diseases associated with bronchoconstriction. Therefore, pulmonary absorption by bronchial tissue determines its local efficacy and was thus investigated in a diffusion cell in vitro. Bronchial epithelium was equilibrated in PBS and discs of 4 mm2 were mounted on that diffusion cell separating the donor and receiver compartment. The donor compartment contained the drug dissolved in PBS (1 mg/ml) and the receiving chamber was permanently flushed with a low flow (1.5 ml/h) of PBS thus allowing time-resolved fractionation for subsequent direct analysis by LC-ESI MS/MS in MRM mode. Transition to the product ion at m z 124 was monitored for quantification (Table 9). The transfer of ipratropium was characterized by the flux (about 220 ng/cm2/min) and the permeability coefficient calculated to be 1.6 x 10-8 cm/s. 

The in vitro methods used to study the percutaneous absorption of drugs vary in the types of diffusion cells used, the skin sources used, and the techniques used to prepare skin for in vitro studies (1). We have attempted to improve the reproducibility and efficiency of measuring the in vitro flux rates of compounds through human skin when carrying out these experiments on a large number of diffusion cells. 

The above procedure was also employed to investigate buccal absorption from the HEMAC experimental delivery device. As in the case of the diffusion cell the drug-loaded disc was positioned on the inner central surface of the buccal mucosa. An impermeable film coated with mucosal adhesive (F-4000, Adhesives Research, Glen Rock, PA) on the periphery was then positioned over the HEMAC disc to prevent dehydration and to secure the device in place on the mucosal surface. The disc was allowed to remain in contact with the mucosa for 4 h before it was removed for quantitation of residual drug content. Blood samples were collected over the same interval as for the saturated solution and processed in the same manner. 

L-a-methyldopa a substrate for the amino acid transporter. In Caco-2 cells, the active transport of this dmg by the amino acid transporter was seven times higher than transport by passive diffusion. Its absorption may be further increased by upregulating the amino acid transporter, as has been observed in the 20-70% stimulation of carrier-mediated amino acid transport by treatment of 0.2 mg/kg growth hormone. 

Bronaugh, R.L. and R.F. Stewart (1984). Methods for in vitro percutaneous absorption studies IV The flow-through diffusion cell, J. Pharmacol Scl, 74, 64-67. 

Clowes, H.M., R.C. Scott and J.R. Heylings (1994). Skin absorption flow-through or static diffusion cells, Toxicol. In Vitro, 8, 827-830. 

Theoretically, a lipophilic drug may pass through the cell or go around it. If drug has a low molecular weight and is lipophilic, the lipid cell membrane is not a barrier to drug diffusion and absorption. In the intestine, molecules smaller than 500 MW may be absorbed by... 

For assessing the absorption potential of specific drug candidates or conducting studies evaluating correlations between drug structure and transport, in vitro models may provide the best approach. Both the tissue-diffusion cell systems and cultured colon cell lines have proved to be particularly useful. 

The biomembrane passage of a drug depends primarily on its physicochemical properties and especially on its partition coefficient (Chapters 22 and 34). Thus, the transient attachment of a lipophilic carrier group to an active principle can provide a better bioavailability, mostly by facifitating cell membrane crossing by passive diffusion. Peroral absorption, as well as rectal absorption, ocular drug delivery and dermal drug delivery, are dependent on passive diffusion. Finally, lipophilic carriers can sometimes be useful to reduce first-pass metabolism. ... 

When bruise causes damage to the internal structure of the apple flesh, the cells are destroyed and the cytoplasm invades the intercellular space this leads to a change in diffusion and absorption coefficients, creating easily detected black spots. 

To estimate percutaneous lethal doses, Dugard and Mawdsley (1978) measured CN transport across human epidermis using a diffusion cell technique, and the data obtained allowed absorption for differing conditions for example, 10% NaCN at pH 11.4 in contact with skin may lead to symptoms within 25 min and death within 1 h. 

With the difficulties associated with accurate estimation of permeability based only on physicochemical properties, a variety of methods of measuring permeability have been developed and used, among which are (l)cul-tured monolayer cell systems, such as Caco-2 or MDCK ( 2 diffusion cell systems that use small sections of intestinal mucosa between two chambers (3) in situ intestinal perfusion experiments performed in anesthetized animals such as rats and (4)intestinal perfusion studies performed in humans (40,54-62). All of these methods offer opportunities to study transport of drug across biological membranes under well-controlledconditions. Caco-2 mono-layer systems in particular have become increasingly commonly used in recent years and human intestinal perfusion methods are also becoming more commonly available. Correlations between Caco-2 permeability and absorption in humans have been developed in several laboratories (63-72). As shown in Fig. 

The receptor phase of any diffusion cell should provide an accurate simulation of the in vivo permeation conditions. The permeant concentration in the receptor fluid should not exceed 10 percent of saturation solubility (Skelly et al. 1987), as excessive receptor phase concentration can lead to a decrease in absorption rate and result in an underestimate of bioavailability. The most common receptor fluid is pH 7.4 phosphate-buffered saline (PBS), although if a compound has a water solubility below 10 p.g/mL, then a wholly aqueous receptor phase is unsuitable, and addition of solubilizers becomes necessary (Bronaugh 1985). 

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