A displacement approach for competitive binding studies was developed. The method utilizes the potentiometric 1-anilino-8-naphthalene-sulfonate (ANS) probe technique and is applied to the binding study of several non-steroidal anti-inflammatory drugs (NSADs) to bovine serum albumin (BSA). A home-made ANS electrode was used to monitor the displaced free ANS probe from its binding sites on the protein molecule by the stepwise addition of the studied drug. To assess and compare quantitatively the displacing ability of the various drugs, the `ANS Displacement Index' is used. The possible interference of 19 ionizable drugs (NSADs, sulfonamides, etc.) to the ANS selective electrode at pH 7.4 was studied and their potentiometric selectivity coefficients (K-ANS,D(pot)) were determined. Correction procedures for the determination of the free ANS concentration are proposed in the case of interfering ionic drugs. A blank binding experiment in conjunction with the incorporation of K-ANS,D(pot) values in the `general competitive site oriented model' allows one to derive estimates for the drug binding parameters, i.e. the number of binding sites and association constants. (C) 1999 Elsevier Science B.V. All rights reserved.

Purpose. To develop an approach based on computer simulations for the study of intestinal drug absorption. Methods. The drug flow in the gastrointestinal tract was simulated with a biased random walk model in the heterogeneous tube model (Pharm. Res. 16, 87-91, 1999), while probability concepts were used to describe the dissolution and absorption processes. An amount of drug was placed into the input end of the tube and allowed to flow, dissolve and absorb along the tube. Various drugs with a diversity in dissolution and permeability characteristics were considered. The fraction of dose absorbed (F-abs) was monitored as a function of time measured in Monte Carlo steps (MCS). The absorption number An was calculated from the mean intestinal transit time and the absorption rate constant adhering to each of the drugs examined. Results. A correspondence between the probability factor used to simulate drug absorption and the conventional absorption rate constant derived from the analysis of data was established. For freely soluble drugs, the estimates for F-abs derived from simulations using as an intestinal transit time 24500 MCS (equivalent to 4.5 h) were in accord with the corresponding data obtained from literature. For sparingly soluble drugs, a comparison of the normalized concentration profiles in the tube derived from the heterogeneous tube model and the classical macroscopic mass balance approach enabled the estimation of the dissolution probability factor for five drugs examined. The prediction of F-abs can be accomplished using estimates for the absorption and the dissolution probability factors. Conclusions. A fully computerized approach which describes the flow, dissolution and absorption of drug in the gastrointestinal tract in terms of probability concepts was developed. This approach can be used to predict F-abs for drugs with various solubility and permeability characteristics provided that probability factors for dissolution and absorption are available.

Phase plane plots are graphical expressions for differential equations ploting the state derivative dc/dt versus the state c. Using these plots, we developed a novel method for the estimation of the terminal slope from time-concentration data. The values of the derivatives used for the construction of the phase plane plots were calculated by two different methods of numerical differentiation. The first method (D1) is based on the classical calculation of slope of the line connecting two successive data points. The alternative method (D2) relies on an initial second-order polynomial interpolation utilizing three successive data points followed by the calculation of the derivative at each one of the concentration values. A forced-through-zero linear regression of the phase plane plot data is used to derive an estimate for the slope. For comparative purposes, the standard approach based on the semilogarithmic plot was also applied. For a hypothetical drug absorbed by first-order process into a one-compartment model, simulated time-concentration data disturbed by a Gaussian zero mean random error with various coefficients of variation were generated. Various sampling schedules, with two, three, four, or five data points, were utilized for the estimation of the terminal slope. Performances of the proposed methods on simulated data were expressed by means of root-mean-squared error, bias, and standard deviation. In all cases, D2 was superior to D1. The D2 method outperforms the standard method in that it furnishes estimates closer to the real values in all cases when two data points and in most cases when three data points were used. All methods behave similarly when four or five data points were used.

A recursion equation which relies on the population growth model of dissolution is used for the analysis of supersaturated dissolution data. The concentration-time data of dissolution experiments are initially transformed to fractions of dose dissolved-generations by adopting an appropriate time interval as the time step of the recursion equation. A computer program is used to derive estimates for the maximum fraction of dose dissolved and the fraction of dose remaining in solution at steady state. Good fittings were observed when this equation was applied to phenytoin and nifedipine supersaturated dissolution data obtained from literature. (C) 1999 Elsevier Science B.V. All rights reserved.

The interaction of diflunisal ion (DF) with beta-cyclodextrin (PCD), gamma-cyclodextrin (gamma CD), and hydroxypropyl-beta-cyclodextrin (HP beta CD) was studied in phosphate buffer, pH 7.4, at 5-37 degrees C and various CD concentrations using a home-made diflunisal ion-selective electrode. Typical direct binding plots and Scatchard plots were obtained with HP beta CD. The Scatchard model for one class of binding sites was used for the estimation of binding parameters for the DF/HP beta CD interaction. The estimates for n (number of binding sites per CD molecule) were in all cases very close to unity, indicating 1:1 complexation. The association constant (K) estimates decrease with increasing temperature. Sigmoidal direct binding plots and concave-downwards Scatchard plots were obtained with various beta CD or gamma CD concentrations. The Hill model was used for the estimation of the binding parameters for the DF/beta CD and DF/gamma CD interactions. Both the Hill coefficients and the binding constants were markedly dependent on the CD concentration. These findings indicate the cooperative character of DF/beta CD and DF/gamma CD interactions. The foe energy change, Delta G, and the thermodynamic parameters, Delta H and Delta S, were estimated for each of the interactions studied using the Van't Hoff equation. (C) 1999 Published by Elsevier Science B.V. All rights reserved.

An isocratic reversed-phase HPLC method was developed to determine cefepime levels in plasma and vitreous fluid. Cefepime and the internal standard cefadroxil were separated on a Shandon Hypersil BDS C18 column by using a mobile phase of 25 mM sodium dihydrogen phosphate monohydrate (pH 3) and methanol (87:13, v/v). Ultraviolet detection was carried out at 270 nm. The retention times were 4.80 min for cefepime and 7.70 min for cefadroxil. This fast procedure which involves an efficient protein precipitation step (addition of HClO4), allows a quantification limit of 2.52 mu g ml(-1) and a detection limit of 0.83 mu g ml(-1). Recoveries and absolute recoveries of cefepime from plasma were 96.13-99.44% and 94-102.5% respectively. The intra-day and inter-day reproducibilities were less than 2% for cefepime at 10, 30, 50 mu g ml(-1) (n=10). The method was proved to be suitable for determining cefepime levels in human plasma and was modified to measure vitreous fluid samples. (C) 1999 Elsevier Science B.V. All rights reserved.

Purpose. A Monte-Carlo computer simulation technique was employed to study the details of the small intestinal transit flow in the gastrointestinal (GI) tract. Methods. A heterogeneous tube model was constructed using a numerical computer simulation technique. The model was built from first principles and included several heterogeneous characteristics of the GI tract structure. We used a random, dendritic-type internal structure representing the villi of the GI tract. The small intestinal transit flow was simulated using two diffusion models, namely, the blind ant and the myopic ant models, which are different models to account the elapse of time, and which are both based on statistical properties of random walks. For each one of the models we utilize two types of biased random walk, placing different emphasis in the motion towards the output of the tube. We monitored the flow of the drug in terms of Monte-Carlo time steps (MCS) through the tube walls and dendritic villi present. Results. The frequency of the transit times was dependent on the structure of the dendritic villi and on the type of biased random walk. The small intestinal flow profile of literature data for a large number of drugs was well characterized by the heterogeneous model using, as parameters, a certain number of villi per unit length of the tube and specific characteristics for both types of the biased random walk. A correspondence between the MCS and real time units was achieved. Conclusions. The transit process of the oral dosage forms in the GI tract can be reproduced with the heterogeneous model developed. This model can be used to study GI absorption phenomena.