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PubMed2003

Comparison of microdialysis and push-pull perfusion for retrieval of serotonin and norepinephrine in the spinal cord dorsal horn.

Both push-pull and microdialysis methods are utilized to measure norepinephrine and serotonin in the dorsal horn of the spinal cord. This experiment was designed to determine which technique is better for measurement of norepinephrine and serotonin in the spinal cord and also to determine if the samples are best collected with or without perchloric acid. Sample stability and an assay validation for precision, limit of quantification, and limit of detection were also performed. Push-pull or microdialysis catheters were placed transversely through the dorsal horn and the catheter was perfused with artificial cerebrospinal fluid. Noxious pinch (20 s/min for 10 min) was used to evoke a change in the concentration of catecholamines. Samples were collected before, during and after pinch. No basal concentrations of epinephrine and serotonin were found with microdialysis. Although basal concentrations of norepinephrine were measured by microdialysis, there was no change in response to noxious pinch. The push-pull technique coupled with collection of samples without perchloric acid showed that significant increases in serotonin and norepinephrine are measurable in response to noxious pinch. In contrast, when samples were collected with perchloric acid present there was no change in serotonin or norepinephrine in response to pinch. The stability of catecholamines is greatly affected by perchloric acid such that there is a near complete loss of ability to detect serotonin and norepinephrine by 24 h in samples collected by push-pull. In contrast, samples collected without perchloric acid showed only a 20% reduction in concentration by 24 h. Even without perchloric acid, by 1 wk there was a 50% or greater loss in the concentrations of norepinephrine in push-pull samples. Thus, to measure changes in catecholamines in the dorsal horn, push-pull collected without perchloric provides measurable, reliable and valid results if analyzed by high performance liquid chromatography within 24 h.

Animals↗

PubMed1996

Effect of brain microdialysis on aminergic transmitter levels in repeated cerebral global transient ischemia in rat.

The effect of repeated transient global ischemia and microdialysis on changes in aminergic neurotransmitter release was investigated using the rat four-vessel occlusion model of global ischemia. To examine the possible transient or permanent changes in neurotransmitter release, ischemia was induced at varying time points in 5 groups of rats. The first ischemia occurred either 24 h (groups I, II, IV, V) or 96 h (group III) following vertebral artery electro-coagulation and guide probe implantation(s), and the second ischemia was induced either 48 h (groups I, IV, V) or 72 h (group II) following the first ischemia. To assess the consequence of repeated microdialysis on the results, one group of rats (group IV) was not dialysed during the first ischemia and another group (group V) was bilaterally dialysed during the second ischemia. Finally, amphetamine-induced neurotransmitter release was also studied in rats submitted to ischemia and compared with that in normal rats. In each case, dopamine, serotonin and their main metabolites were measured by HPLC with electrochemical detection. Monoamine release was inhibited during the second episode of transient ischemia; this non-release was linked to the repeated microdialysis and not to the repeated ischemia. Although the results of chronic studies using brain microdialysis have been widely recognized as valid, the findings presented here indicate that combined with ischemia, probe reinsertion modifies the level of neurotransmitter release.

Amphetamines↗

PubMed1998

Neurochemical and morphological responses to acutely and chronically implanted brain microdialysis probes.

The purpose of this study was to compare, in rats, brain microdialysis results obtained using microdialysis probes implanted acutely for 2 h versus probes implanted chronically for 24 h in the caudate. Specific comparisons included: (1) dialysate purine and amino acid profiles during cerebral ischemia; (2) diffusional characteristics of the microdialysis probe; and (3) tissue morphology surrounding the probe. During ischemia, the increase in dialysate levels of adenosine, inosine, and hypoxanthine was less pronounced from probes implanted chronically, while dialysate xanthine levels increased to a greater extent. An increase in dialysate amino acid neurotransmitters during cerebral ischemia was observed in the acutely implanted probes within 10 min of the onset of cerebral ischemia; in the chronically implanted probes this increase did not occur until after 50 min of severe ischemia. Both in vitro and in vivo tests revealed a diffusional barrier in chronically implanted probes. Moreover, the tissue surrounding chronically implanted probes exhibited a high degree of inflammation, and fibrin deposits were substantial. In addition, uric acid levels (an indicator of tissue injury) sampled from chronically implanted probes were 7-fold greater than levels sampled from acutely implanted probes. These data raise concerns about the use of chronically implanted microdialysis probes for the measurement of purine and amino acid profiles during cerebral ischemia.

Adenosine↗

PubMed2000

Direct interaction between the sympathetic and renin-angiotensin system in myocardial tissue: a microdialysis study in anaesthetised rats.

It has been suggested that local activation of the renin-angiotensin system is involved in early stages of myocardial pathophysiology. To date, there is increasing evidence for interactions between the renin-angiotensin system and the sympathetic nervous system; consequently, local sympathetic activation may also be involved in this. Microdialysis has great potential in the direct investigation of neurohormonal interactions. Therefore, the present study employs microdialysis to study the local effects of exogenous angiotensin II on the interstitial norepinephrine concentration of the normally innervated left ventricle of the anaesthetised rat. The present study investigates the effect of increasing dosages of exogenous angiotensin II on local interstitial norepinephrine. Furthermore, a single dose of losartan was infused on top of the highest dose of angiotensin II, in order to study possible involvement of angiotensin II type 1 (AT1) receptors. Both infusion and sampling were carried out locally, via the microdialysis probes. Concomitantly, circulating norepinephrine levels, heart rate and respiratory rate were monitored to evaluate physiologic stability of the preparation throughout the experiment. Time controls consisted of rats that were perfused with only a Ringer's solution. Angiotensin II induced a dose dependent increase in norepinephrine that was significantly reduced by losartan. Norepinephrine levels in both plasma (infusion experiment and time controls) and the left ventricular wall (time controls) remained stable throughout the experiment, just as heart rate and respiratory rate did. This study for the first time employs microdialysis to demonstrate direct interaction between the sympathetic nervous system and the renin-angiotensin system in the rat left ventricle. The data strongly suggest that AT1 receptors are involved in this interaction, since selective AT1 receptor blockade with losartan significantly reduced the angiotensin II induced norepinephrine concentration.

Angiotensin II↗

PubMed2000

Intradermal microdialysis: kinetics of iontophoretically delivered propranolol in forearm dermis.

Intradermal microdialysis permits us to measure the concentration in dermis of drugs applied to the skin. Microdialysis is especially efficient in sampling water-soluble molecules. Consequently, it appears particularly suitable to study current based delivery systems like iontophoresis that deliver ions or highly polar molecules. The purpose of this work was to evaluate the adequacy of a skin microdialysis technique to characterize and quantify the dermatopharmacokinetics of iontophoretically delivered propranolol in the dermis of healthy human volunteers. Linear microdialysis probes were inserted in the subject's forearm skin and an iontophoresis device was installed above them. Constant current was applied for two periods of 1 h each separated by a 1-h interval. Dialysate samples were collected every 6 min for 4.4 h and analyzed by HPLC. Probes were always placed in the dermis as measured by ultrasonography. Propranolol was detectable in the dialysate. It was possible to build detailed concentration vs. midtime profiles that mirrored the current applied. Elimination rate from the dermis had first-order kinetics and was similar in all subjects. Quantification of the absorption process, indexed by lag-time and area under the concentration curve showed a high inter- and intrasubject variability that did not correlate with probe depth.

Adult↗

PubMed2000

Application of microdialysis to characterize drug disposition in tumors.

Microdialysis is an in vivo sampling technique that was initially developed to measure endogenous substances in the field of neurotransmitter research. In the past decade, microdialysis has been increasingly applied to study the pharmacokinetics and drug metabolism in the blood and various tissues of both animals and humans. This paper describes the general aspects of this in vivo sampling technique followed by the survey of the recent papers regarding the application of microdialysis to characterize anticancer drug disposition in solid tumors. It can be concluded that microdialysis is a very suitable method to obtain drug concentration-time profiles in the interstitial fluid of solid tumors as well as of other variety of tissues.

Animals↗

PubMed1998

In vivo microdialysis reveals facilitatory metabotropic glutamate receptors regulating excitatory amino acid release in rat nucleus tractus solitarius.

The functional involvement of metabotropic glutamate receptors in nucleus tractus solitarius of the medulla has been examined by evaluating the action of the selective metabotropic glutamate receptor agonist trans-1-amino-cyclopentane-1,3-dicarboxylate on the extracellular release of L-glutamate and L-aspartate by in vivo microdialysis. Studies were performed in urethane anaesthetized rats, with a microdialysis probe being inserted into the medial nucleus tractus solitarius, which was constantly perfused with artificial cerebrospinal fluid (1 microliter/min). Following an equilibration period, samples were collected for amino acid analysis by HPLC. trans-1-Amino-cyclopentane-1,3-dicarboxylate (30 microM) produced 190% and 500% increases in basal release of L-glutamate and L-aspartate, respectively. trans-1-Amino-cyclopentane-1,3-dicarboxylate-evoked release of L-glutamate and L-aspartate was concentration-dependent (10-100 microM), and was calcium and tetrodotoxin-sensitive. Two metabotropic glutamate receptor antagonists, (+)-alpha-methyl-4-carboxyphenylglycine (200 microM) and (S)-4-carboxyphenylglycine (500 microM), when administered via a microdialysis probe, significantly attenuated the trans-1-amino-cyclopentane-1,3-dicarboxylate-evoked release of both L-glutamate and L-aspartate. Basal release was not altered by these two antagonists. Histological studies verified the location of the probe in the nucleus tractus solitarius. These studies provide the first evidence using in vivo microdialysis of functional metabotropic glutamate receptors, which seem to be facilitatory pre- and postsynaptically located receptors, in the rat nucleus tractus solitarius. On the basis of our findings, metabotropic glutamate receptors are likely to play key modulatory roles in regulating transmitter release.

Animals↗

PubMed2003

Intestinal ischaemia during cardiac arrest and resuscitation: comparative analysis of extracellular metabolites by microdialysis.

Intestinal ischaemia is a major complication of shock syndromes causing translocation of bacteria and endotoxins and multiple organ failure in intensive care patients. The present study was designed to use microdialysis as a tool to monitor intestinal ischaemia after cardiac arrest and resuscitation in pigs. For this purpose, microdialysis probes were implanted in pig jejunal wall, peritoneum, skeletal muscle and brain, and interstitial fluid was obtained during circulatory arrest (induced by ventricular fibrillation) and after return of spontaneous circulation (ROSC). Cardiac arrest for 4 min caused a prolonged (60 min) reduction of blood flow in jejunal wall, muscle and brain as determined by the ethanol technique. This was accompanied by cellular damage in heart muscle and brain as indicated by increased levels of troponin-I and protein S-100, respectively. Plasma levels of glucose, lactate and choline were increased at 15-60 min following cardiac arrest. In contrast, cardiac arrest induced a rapid but variable decrease of interstitial glucose levels in all monitored organs; this decrease was followed by an increase over baseline during reperfusion. In the intestine, lactate, glutamate and choline levels were increased during ischaemia and reperfusion for 60-120 min; intestinal and peritoneal samples yielded parallel changes of lactate levels. Brain and muscle samples showed similar changes as in intestinum and peritoneum except for glutamate, which was increased in brain but not in muscle. We conclude that intestinal ischaemia occurs as a consequence of cardiac arrest and resuscitation and can be monitored by in vivo microdialysis. Comparative analysis by multi-site microdialysis reveals that the intestine is equally or even more sensitive to ischaemia than brain or muscle.

Animals↗

PubMed2003

Brain metabolism during cardiopulmonary resuscitation assessed with microdialysis.

BACKGROUND AND PURPOSE: Microdialysis is an established tool to analyse tissue biochemistry, but the value of this technique to monitor cardiopulmonary resuscitation (CPR) effects on cerebral metabolism is unknown. The purpose of this study was to assess the effects of active-compression-decompression (ACD) CPR in combination with an inspiratory threshold valve (ITV) (=experimental CPR) vs. standard CPR on cerebral metabolism measured with microdialysis. METHODS: Fourteen domestic pigs were surfaced-cooled to a body core temperature of 26 degrees C and ventricular fibrillation was induced, followed by 10 min of untreated cardiac arrest; and subsequently, standard (n=7) CPR vs. experimental (n=7) CPR. After 8 min of CPR, all animals received 0.4 U/kg vasopressin IV, and CPR was maintained for an additional 10 min in each group; defibrillation was attempted after a total of 28 min of cardiac arrest, including 18 min of CPR. RESULTS: In the standard CPR group, microdialysis measurements showed a 13-fold increase of the lactate-pyruvate ratio from 7.2+/-1.3 to 95.5+/-15.4 until the end of CPR (P<0.01), followed by a further increase up to 138+/-32 during the postresuscitation period. The experimental group developed a sixfold increase of the lactate-pyruvate ratio from 7.1+/-2.0 to 51.1+/-8.7 (P<0.05), and a continuous decrease after vasopressin. In the standard resuscitated group, but not during experimental CPR, a significant increase of cerebral glucose levels from 0.6+/-0.1 to 2.6+/-0.5 mM was measured (P<0.01). CONCLUSION: Using the technique of microdialysis we were able to measure changes of brain biochemistry during and after the very special situation of hypothermic cardiopulmonary arrest. Experimental CPR improved the lactate-pyruvate ratio, and glucose metabolism.

Analysis of Variance↗

PubMed2001

L-ephedrine-induced neurodegeneration in the parietal cortex and thalamus of the rat is dependent on hyperthermia and can be altered by the process of in vivo brain microdialysis.

Multiple doses of the dietary supplement L-ephedrine can cause severe hyperthermia and modest dopamine depletions in the rat brain. Since D-amphetamine treatment can result in neurodegeneration, the potential of L-ephedrine to produce similar types of degeneration was investigated. Adult male rats, some implanted in the caudate/putamen (CPu) for microdialysis, were given four doses of 25 mg/kg L-ephedrine or 5 mg/kg D-amphetamine (2 h between doses) at an ambient temperature of 23 degrees C. L-ephedrine-induced degeneration in the forebrain was dependent on the degree of hyperthermia. Layer IV of the parietal cortex was the most sensitive to L-ephedrine treatment with peak body temperatures of at most 40.0 degrees C necessary to produce degeneration. Extensive neurodegeneration in the parietal cortex after L-ephedrine treatment was as pronounced as that previously described for D-amphetamine treatment and also occurred in the intralaminar, ventromedial and ventrolateral thalamic nuclei in rats with severe hyperthermia (peak body temperatures>41.0 degrees C). The neurodegeneration induced by L-ephedrine may have resulted in part from excitotoxic mechanisms involving the indirect pathways of the basal ganglia and related areas. No differences were observed between microdialysis and non-implanted rats with respect to degree of tyrosine hydroxylase (TH) loss in the CPu after either D-amphetamine or L-ephedrine treatment. However, neurodegeneration resulting from D-amphetamine and L-ephedrine was reduced in the microdialysis animals in the hemisphere ipsilateral to the probe, which raises concerns when using the technique of in vivo microdialysis to evaluate neurodegeneration. The results of this study, in conjunction with human clinical evaluation of ephedrine neurotoxicity, indicate that regionally specific damage may occur in the cortex of some humans exposed to ephedrine in the absence of stroke or hemorrhage.

Animals↗

PubMed1997

Capillary electrophoresis and microdialysis: current technology and applications.

Microdialysis sampling has become an important method for the continuous monitoring from an in vivo environment. This technique has been used to monitor many endogenous molecules, such as neurotransmitters, as well as exogenous species such as drug substances. Microdialysis samples have traditionally been analyzed by liquid chromatographic (LC) methods to gain resolution and quantification of the molecules of interest. However, LC separations have a relatively large injection volume requirement which, as a consequence, increases microdialysis sampling times. Capillary electrophoresis (CE), with its very small sample volume requirements and high resolving power, has therefore gained popularity as an alternative to LC. Reviewed here are many of the technologies currently available for CE and examples of how this technique has been effectively applied to the analysis of microdialysis samples.

Amino Acids↗

PubMed1998

Determination of glutamate and aspartate in microdialysis samples by reversed-phase column liquid chromatography with fluorescence and electrochemical detection.

Five different systems for fast determination of aspartate and glutamate in microdialysis samples are described: (I) a high-speed HPLC using a gradient pump with a sharp elution profile, (II) a column switching technique, (III) an isocratic pump with a low-pressure switching valve for one-step gradients, (IV) microbore chromatography using injections of acetonitrile as a wash-out step, (V) on-line connection of microdialysis and HPLC/derivatization. In all cases, automated precolumn derivatization with o-phthalaldehyde-2-mercaptoethanol reagent were used. Both fluorescence and electrochemical detection techniques were evaluated in terms of reproducibility, sensitivity, interference, maintenance and troubleshooting. The electrochemical detection method required a second derivatization step with 0.2 M iodoacetamide to remove excess of a thiol moiety and regular recalibrations after each six to ten injections. Under these conditions the correlation coefficients for electrochemical vs. fluorescence detectors were 0.918 for Asp and 0.988 for Glu for 65 microdialysis samples. Coefficients of variation for six analyses between calibrations were below 3% for both detectors. The limits of detection for both amino acids were about 0.4 pmol for electrochemical detection with a thiol scavenger step, 50 fmol for fluorescence detection using conventional columns and about 20-30 fmol for the microbore system. All systems are suitable for detecting basal levels of Asp and Glu in 5-10 microl microdialysis samples from a rat brain where typical concentrations lie around 1-10 pmol or more. It is concluded that a microbore setup with one isocratic pump and an autosampler optimized for injections of washing solvent between samples is the most practical and economical. The system allows analysis of minute sample volumes down to 1-2 microl.

Animals↗

PubMed1999

Measurement of unbound caffeic acid in rat blood by on-line microdialysis coupled with liquid chromatography and its application to pharmacokinetic study.

To monitor the levels of caffeic acid in rat blood, an on-line microdialysis system coupled with liquid chromatography was developed. The microdialysis probe was inserted into the jugular vein/right atrium of male Sprague-Dawley rats. Caffeic acid (100 mg/kg, i.v.) was then administered via the femoral vein. Dialysates were automatically injected onto a liquid chromatographic system via an on-line injector. Samples were eluted with a mobile phase containing methanol-100 mM monosodium phosphoric acid (35:65, v/v, pH 2.5). The UV detector wavelength was set at 320 nm. The detection limit of caffeic acid was 20 ng/ml. The in vivo recoveries of the microdialysis probe for caffeic acid at 0.5 and 1 microg/ml were 48.34+/-2.68 and 47.64+/-3.43%, respectively (n=6). Intra- and inter-assay accuracy and precision of the analyses were < or = 10% in the range of 0.05 to 10 microg/ml. Pharmacokinetics analysis of results obtained using such a microdialysis-chromatographic method indicated that unbound caffeic acid in the rat fitted best to a biexponential decay model.

Animals↗

PubMed1999

Sensitive and specific method for the simultaneous determination of natural and synthetic catecholamines and 3,4-dihydroxyphenylglycol in microdialysis samples.

The relatively new technique of microdialysis provides new possibilities for investigating in vivo the functioning of the sympathetic nervous system. The small sample volumes obtained, however, are a great challenge for analytical chemists. We report here a HPLC method for measuring in one run both natural and synthetic catecholamines [dopamine, (nor)epinephrine, alpha-methylnorepinephrine, isoproterenol and epinine] and the intraneuronal metabolite 3,4-dihydroxyphenylglycol in small microdialysis samples after derivatization with the fluorogenic agent 1,2-diphenylethylenediamine. No prior clean-up step is necessary. N-Ethylmaleimide is necessary for preventing an inhibitory action on derivatization occurring in in vivo microdialysis samples. The method can handle large numbers of samples, is sensitive (on-column detection limits 30 to 200 fg) and reproducible (RSD 1 to 7%). Recovery characteristics of the commercial microdialysis probe used (CMA/20) were extensively investigated both in vitro and in vivo at various perfusion rates; for practical purposes a rate of 2 microl/min and sampling at 10-min intervals was found to be workable and to give good and reproducible recoveries (50 to 70%).

Animals↗

PubMed1999

Evaluation of transdermal iontophoresis of enoxacin from polymer formulations: in vitro skin permeation and in vivo microdialysis using Wistar rat as an animal model.

Polymers were used in vehicles to form hydrogel matrices in this study to evaluate the in vitro permeation and in vivo microdialysis of enoxacin. The highest transdermal delivery determined by area under flux-time curve (AUC) and intracutaneous enoxacin concentration were observed in methylcellulose (MC) and polyvinylpyrrolidone (PVP) hydrogels, respectively. To avoid the pH shift in vehicles during iontophoresis, buffer species were added to formulations to increase the buffer capacity. As expected, the permeability of enoxacin of anodal iontophoresis was larger than that of cathodal iontophoresis. Combination of benzalkonium chloride, a cationic surfactant as an enhancer, and iontophoresis exerted an enhancing effect for anionic enoxacin at pH 10.0. However, no effect or a negative effect was detected for cationic enoxacin in deionized water or pH 5.0 buffer, due to the shielding of the negative charge in the skin. The skin residue of enoxacin was slightly increased after the incorporation of Azone in PVP hydrogel. The result of in vivo microdialysis was in accordance with that of in vitro study. The effect of Azone on the intracutaneous enoxacin was more significant for in vivo microdialysis than in the in vitro study indicating the clinical feasibility of Azone for iontophoretic delivery. Microdialysis can be considered as a useful technique to investigate the pharmacokinetics of transdermal iontophoresis in vivo.

Administration, Cutaneous↗

PubMed2001

Determination of myocardial norepinephrine in freely moving rats using in vivo microdialysis sampling and liquid chromatography with dual-electrode amperometric detection.

Myocardial norepinephrine (NE) is considered a meaningful parameter for estimation of cardiac function. Long lasting changes in myocardial NE appear to be not only a consequence of pathologic processes in the myocardium, but may be a factor responsible for some diseases (e.g. increased propensity for arrhythmias or negative effect on left ventricular contractility in congestive heart failure). In this respect monitoring of myocardial NE is of great importance. A microdialysis sampling technique coupled with liquid chromatography with electrochemical detection (LCEC) was developed to measure the in vivo NE concentration in the myocardial interstitium of conscious, freely moving rats. LCEC using a dual-electrode amperometric detection in the series configuration provided detection limits for NE of 10 pg/ml in 20 microl microdialysis samples. Microdialysis probes of the linear design were implanted in the myocardial tissue in the periphery of the left descending coronary artery. The basal steady-state concentration of NE in myocardial dialysate of awake, freely moving rats was found to be 0.17+/-0.026 ng/ml. Delivery through the microdialysis probe of the NE reuptake inhibitor desipramine (DMI) at a concentration of 0.1 mM increased NE release to 153+/-13% of control. If the concentration of DMI in the perfusate was increased to 1.0 mM, NE release increased to only 166+/-21% of control.

Adrenergic Uptake Inhibitors↗

PubMed2003

Improving microdialysis extraction efficiency of lipophilic eicosanoids.

Microdialysis recovery of the lipophilic analytes prostaglandin B2, leukotriene B4 and C4 was studied in vitro. Relative recovery (RR) through different commercially-available microdialysis probes for prostaglandin B2 and leukotrienes was examined using different flow rates. The enhancing effect at different concentrations of binding agents such as alpha, beta, gamma-cyclodextrins (alpha, beta, gamma-CD) on the microdialysis RR for different eicosanoids was evaluated. Small organic molecules such as ethanol, propylene glycol and dimethyl sulfoxide (DMSO) were studied in terms of their effect on enhancing RR. Inclusion of arachidonic acid in either the perfusion fluid or the sample medium caused the microdialysis RR for these hydrophobic analytes to be increased.

Chromatography, High Pressure Liquid↗

PubMed2003

Flow-injection chemiluminescence detection for studying protein binding of terbutaline sulfate with on-line microdialysis sampling.

The binding of terbutaline sulfate to bovine serum albumin was studied in vitro using the technique of microdialysis sampling combined with flow-injection chemiluminescence analysis (FIA-CL). In the presence of formaldehyde, terbutaline sulfate can be oxidized by KMnO(4) to produce high chemiluminescence emission in sulfate acid media. The concentration of terbutaline sulfate is proportional with the CL intensity in the range of 1 x 10(-7)-2 x 10(-5) mol l(-1) with a detection limit of 3 x 10(-8) mol l(-1). The drug and protein were mixed in different molar ratios in 0.067 mol l(-1) phosphate buffer, pH 7.4, and incubated at 37 degrees C in a water bath. The microdialysis probe was utilized to sample the mixed solution at a perfusion rate of 5 microl min(-1) and the dialytic efficiency of terbutaline sulfate under the experimental conditions was 26.3%. The data obtained by proposed microdialysis flow-injection chemiluminescence method was analyzed with Scrathard analysis and Klotz plot. The estimated association constant (K) and the number of the binding site (n) on one molecule of BSA by Scrathard analysis were 4.11 x 10(4) l mol(-1) and 1.06, respectively. The proposed system proved that FIA-CL coupled with on-line microdialysis sampling is a simple and reliable technique for the study of drug-protein interaction.

Animals↗