Penetration of betaxolol HCL ionic suspension 0.25% and betaxolol HCL solution 0.50% into the aqueous humor.

PURPOSE: To determine the intraocular penetration of topical drops of betaxolol HCl 0.25% suspension and betaxolol HCl 0.50% solution into the aqueous humor. METHODS: Fifteen patients were randomly assigned to receive topical betaxolol HCl 0.25% suspension (n=7) or topical betaxolol HCl 0.50% solution (n=8) the day before cataract surgery. Aqueous samples were collected 2 hours after the administration of the morning dose during cataract surgery. Drug concentrations were determined by high-performance liquid chromatography with fluorescence detection. RESULTS: The mean aqueous humor concentration of topical betaxolol HCl 0.25% suspension was 275.1+/-168.8 micro g/mL (range 570-70 micro g/mL) and the mean aqueous humor concentration of topical betaxolol HCl 0.50% solution was 195.4+/-102.4 micro g/mL (range 334-50 micro g/mL) (p=0.281). CONCLUSIONS: The mean aqueous humor concentration of betaxolol 0.25% suspension was higher than betaxolol 0.50% solution; however, the difference was not statistically significant. With twofold reduced concentration and similar anterior chamber penetration, betaxolol 0.25% suspension could be first choice for Beta 1 selective blocker therapy when considered for patients with glaucoma.

Polymorphic debrisoquin metabolism in a Turkish population.

Debrisoquin hydroxylation polymorphism was studied in 326 unrelated healthy Turkish volunteers. Debrisoquin sulfate (10 mg) was administered to subjects, and debrisoquin and 4-hydroxydebrisoquin were determined in the 0- to 8-hour urine samples. Debrisoquin oxidation was polymorphic, with 11 subjects (3.37%; 95% confidence interval, 1.69% to 6.07%) phenotyped as poor metabolizers. The metabolic ratio between debrisoquin and 4-hydroxydebrisoquin in 8-hour urine samples ranged from 0.02 in extensive metabolizers to 263.8 in poor metabolizers. The proportion of poor metabolizers was found to be in the range observed in the other white populations studied.

Subretinal fluid levels of topical, oral, and combined administered ciprofloxacin in humans.

AIMS: To investigate the subretinal fluid (SRF) penetration of ciprofloxacin following topical, oral, and combined administration. METHODS: 34 patients undergoing conventional retinal reattachment surgery were randomly assigned to three groups. Twelve patients received topical ciprofloxacin, 11 patients received oral ciprofloxacin, and the other 11 patients received combined drug administration. SRF drug level was measured by using high performance liquid chromatography method. RESULTS: The highest drug concentrations of all tested modes were attained following combined administration and lowest following topical administration (p<0.001). The SRF drug concentration following oral administration was also significantly higher than that of topical administration (p<0.001). Concentrations after oral and combined administration did not differ significantly (p = 0.217). CONCLUSIONS: Topical ciprofloxacin can penetrate SRF. Ocular bioavailability of ciprofloxacin in SRF after oral and combined administration is equivalent.

Penetration of topical, oral, and combined administered ofloxacin into the subretinal fluid.

AIMS: To assess the subretinal fluid (SRF) levels of ofloxacin following topical, oral or combined administration. METHODS: 31 patients undergoing conventional retinal reattachment surgery were randomly assigned to three groups. Nine patients received topical ofloxacin, 11 patients received oral ofloxacin, and the other 11 patients received combined administration. Collected SRF samples were analysed for drug level by using high performance liquid chromatography. RESULTS: SRF drug levels after oral and combined administration were significantly higher than that after topical administration (p=0.0002 and p=0.0002, respectively) while there was no significant difference between oral and combined administration (p=0.0844). CONCLUSIONS: Ocular bioavailability of ofloxacin in SRF after oral and combined administration is equivalent. The addition of oral ofloxacin to topical therapy increased drug SRF penetration sixfold.

Penetration of topical and oral ofloxacin into the aqueous and vitreous humor of inflamed rabbit eyes.

PURPOSE: This study aimed to investigate the penetration of topical and oral ofloxacin into aqueous humor and vitreous humor in post-traumatic endophthalmitis model in rabbits. METHODS: A standardized intraocular infection after penetrating injury was made in the right eyes of 16 rabbits. Intraocular infection was induced by intravitreal injection of a suspension of Staphylococcus aureus. The intact left eyes were maintained as controls. The animals were divided randomly into two groups. (1) In the topical group, two drops of ofloxacin 0.3% eyedrops were instilled to both eyes every 30 min for 4 h. (2) In the topical-oral group, two doses of 25 mg/kg of ofloxacin at 12-h intervals were given orally, then the protocol of the first group was applied. Aqueous and vitreous humor samples were taken 30 min after the last drop. Ofloxacin concentrations were measured by using HPLC. RESULTS: Mean aqueous levels of ofloxacin in control eyes were: 3.25 +/- 2.55 microg/ml in topical group. 4.58 +/- 5.39 microg/ml in topical-oral group. Mean aqueous levels in inflamed eyes were: 5.21 +/- 4.55 microg/ml in topical group, 10.34 +/- 8.88 microg/ml in topical-oral group. Mean vitreous levels of ofloxacin in control eyes were: 0.17 +/- 0.07 microg/ml in topical group, 1.30 +/- 1.23 microg/ml in topical-oral group. Mean vitreous levels in inflamed eyes were: 0.35 +/- 0.22 microg/ml in topical group, 3.48 +/- 2.69 microg/ml in topical-oral group. There was no significant difference among the groups (P > 0.05), however. CONCLUSIONS: The result of this study suggests that oral supplementation of ofloxacin to topical instillation increased the ocular levels of ofloxacin in the post-traumatic endophthalmitis model. Mean drug concentrations in aqueous and vitreous humors were above the 90% minimum inhibitory concentrations (MIC90) for most of the common microorganisms causing endophthalmitis in all eyes, except in the vitreous humors of the intact eyes instilled topically.

The effects of prolonged acute use and inflammation on the ocular penetration of topical ciprofloxacin.

PURPOSE: To study the aqueous and vitreous penetration of ciprofloxacin after prolonged acute topical administration and to investigate the effects of inflammation on drug penetration. METHODS: A standardized model of intraocular infection after penetrating injury was made in the right eyes of eight rabbits. The intact left eyes were maintained as the control. Two drops of ciprofloxacin 0.3% eyedrops were instilled topically every 1 h for 7 h to all eyes of the rabbits. Aqueous and vitreous samples (100 microl) were obtained half an hour after the last drop. Instillation was continued for 7 h more and samples were obtained as before. Drug concentrations were measured using HPLC. RESULTS: The mean aqueous humor levels of ciprofloxacin were: in control eyes 1.31 +/- 0.78 microg/ml after 7 h and 1.85 +/- 1.69 microg/ml after 14 h of instillation: in inflamed eyes 2.18 +/- 1.02 microg/ml after 7 h and 2.91 +/- 2.12 microg/ml after 14 h. The mean vitreous humor levels were: in control eyes 0.65 +/- 0.44 microg/ml after 7 h and 0.72 +/- 0.8 microg/ml after 14 h of instillation; in inflamed eyes 0.67 +/- 0.77 microg/ml after 7 h and 1.01 +/- 0.43 microg/ml after 14 h. However, the differences among the groups were not significant (P > 0.05). CONCLUSIONS: Ciprofloxacin penetration into aqueous humor was higher in 14-h topical application than that for 7 h. Inflammation increased the penetration of topical ciprofloxacin into aqueous while administered for 7 h and into both aqueous and vitreous humor while administered for 14 h. c

Determination of debrisoquine and 4-hydroxydebrisoquine in urine by high-performance liquid chromatography with fluorescence detection after solid-phase extraction.

A simple, selective and sensitive method has been developed to determine debrisoquine and 4-hydroxydebrisoquine in human urine. Separation of the analytes was obtained using a mobile phase of 0.1 M sodium dihydrogen phosphate-acetonitrile (87:13, v/v) and a muBondapak C18 column. The column effluent was monitored with fluorescence detection at 210 nm (ex) and 290 nm (em). Rapid sample preparation was achieved by solid-phase extraction columns (Bond Elut CBA, 3 ml capacity) which provided excellent recovery values for both compounds. The cost per sample using this approach could be minimized by column regeneration and re-use. The within-day and the day-to-day reproducibilities were less than 7% for both components. The method was shown to be suitable for the study of the debrisoquine-sparteine type genetic polymorphism in man.

Rapid liquid chromatographic assay of ciprofloxacin in human aqueous humor.

A simple, selective and sensitive method has been developed to determine ciprofloxacin in human aqueous humor. Separation of ciprofloxacin was carried out with pipemidic acid as internal standard using a Novapak C18 reversed-phase cartridge column (100 x 8 mm i.d., particle size 4 microns) and a mobile phase consisting of methanol-acetonitrile-citric acid (0.4 M) (3:1:10, v/v/v) at a flow rate of 1 ml min-1. The column effluent was monitored with fluorescence detection at 278 nm (excitation) and 450 nm (emission) after direct injection. The retention times were 4.88 min for pipemidic acid and 7.52 min for ciprofloxacin. The within-day and day-to-day reproducibilities were less than 7% for ciprofloxacin at 0.1 and 1 microgram ml-1 (n = 6). The mean recovery from aqueous humor was found to be 101.37 +/- 6.7% for ciprofloxacin at 0.1 micrograms ml-1 (n = 6 and the detection limit corresponding to a signal-to-noise ratio of 2.5:1 was 250 pg ml-1. The method was shown to be suitable for determining ciprofloxacin levels in human aqueous humor samples.

Optimization of mobile phase in the separation of beta-blockers by HPLC.

Beta-blockers are generally determined using high-performance liquid chromatography (HPLC). Previous HPLC separations of beta-blockers have often required a mobile phase containing three components; acetonitrile or methanol to control the retention; buffer to control the ionic strength and pH of the mobile phase; ion-pairing reagent to provide adequate retention of beta-blockers or organic amines as masking agent to reduce peak tailing. Due to the complexity of the mobile phases employed, development of these assays can be a laborious process. Additionally, alkyl sulphonates and organic amines dramatically reduces the life-time reduction of silica based C18 columns. The results of this study demonstrated that the addition of tested alkyl sulphonates and organic amines is not essential for an adequate separation of beta-blockers. In this study, we developed a simple HPLC method for the simultaneous separation of model beta-blockers, atenolol, practolol, metoprolol, oxprenolol and propranolol. Atenolol, practolol, metoprolol, oxprenolol and propranolol adequately separated with high peak symmetries using a mobile phase consisted of methanol/acetonitrile/phosphate buffer (10 mM, pH 3.0) (15:15:70, v/v/v). By altering only the fraction of methanol with respect to acetonitrile, method development becomes a more efficient separation. Furthermore, atenolol, practolol, metoprolol, oxprenolol and propranolol can be detected up to 0.25, 5, 10, 50 and 10 ng ml(-1). In this publication, we present the simultaneous separation of beta-blockers having a wide range of polarity. It is proposed that this new mobile phase, consisting only acetonitrile, methanol and phosphate buffer can be used for the analysis of the several beta-blockers presently in doping control analysis as well as others.

Determination of ofloxacin in human aqueous humour by high-performance liquid chromatography with fluorescence detection.

A reversed-phase high-performance liquid chromatographic method is described for the determination of ofloxacin in human aqueous humour; the method involves fluorescence detection (excitation at 290 nm; emission at 500 nm) after direct injection of samples. The method utilized a 100 mm x 8 mm i.d. cartridge column packed with 4 microns Novapak C18 with a mobile phase methanol-acetonitrile-0.4 M citric acid (3:1:10, v/v/v) and a flow rate of 1 ml min-1 at ambient temperature. The retention times for the internal standard pipemidic acid and for ofloxacin were 4.82 and 7.32 min respectively. The mean recovery (+/- ISD) from human aqueous humour was 103.24 +/- 4.45% for ofloxacin at 1 microgram ml-1 (n = 6). The within-day and day-to-day RSDs at 0.1 microgram ml-1 and 1 microgram ml-1 were less than 6.71% (n = 6) and the lower limit of reliable determination corresponding to a signal-to-noise ratio of 2.5:1 was 20 ng ml-1. The assay was shown to be suitable for measuring ofloxacin levels in human aqueous humour samples after topical, oral and intravenous administration.

Comparison of aqueous humour and vitreous humour levels of two 0.3% ciprofloxacin eyedrops.

BACKGROUND: Two ophthalmic solutions of 0.3% ciprofloxacin eyedrops are available in Turkey: Ciloxan and Siprogut. The objective of this study was to compare the concentrations of drug produced by the two products in the aqueous humour and vitreous humour after local administration. METHODS: Twenty-one patients undergoing primary vitreoretinal surgery received either Ciloxan (10 patients) or Siprogut (11 patients). Six hours before surgery, two drops of solution were instilled onto the operative eye. Drops were then instilled every 30 minutes for the first 3 hours and then hourly for the next 3 hours. Aqueous and vitreous samples were collected 30 minutes after administration of the last dose and were assayed for ciprofloxacin concentration by means of high-performance liquid chromatography with fluorometric detection. RESULTS: The mean aqueous humour concentrations of Ciloxan and Siprogut were 0.36 (standard error of the mean [SEM] 0.09) microgram/mL and 0.44 (SEM 0.17) microgram/mL respectively. The corresponding vitreous humour concentrations were 0.21 (SEM 0.05) microgram/mL and 0.22 (SEM 0.06) microgram/mL. Neither of these differences was statistically significant. The aqueous and vitreous levels of both products exceeded the minimum inhibitory concentrations for certain bacterial species that frequently cause intraocular infections. INTERPRETATION: Our results show that the ocular bioavailability of Ciloxan and Siprogut after local administration is equivalent. Penetration of ciprofloxacin into the vitreous humour seems to be poorer than that into the aqueous humour.

Metabolic ratios of four probes of CYP2D6 in Turkish subjects: a cross-over study.

The relationships among the metabolic ratios for the standard probe drugs of CYP2D6 activity, such as debrisoquine, sparteine, metoprolol and dextromethorphan, were studied in 32 Turkish subjects. All subjects were randomly selected according to their phenotypes from a group of 111 Turkish subjects whose oxidation status had been tested for debrisoquine previously. All subjects were given a 10 mg debrisoquine tablet, a 100 mg sparteine tablet, a 100 mg. metoprolol tablet and a 20 mg dextromethorphan capsule orally with a wash-out period of at least 1 week between each probe administration. Metabolic ratios were calculated as percentage of dose excreted as parent drug/percentage of dose excreted as its hydroxymetabolite of parent drug in 0-8 h urine. Three poor metabolisers (PM) of debrisoquine were identified. They were also PMs of the other test probes and no misclassification by the 4 phenotyping methods was observed. All six correlations among the metabolic ratios of the 4 probe drugs assessed by Spearman's rank test were highly significant (P < 0.001). The present findings indicate that the oxidative metabolism of debrisoquine, sparteine, metoprolol and dextromethorphan is catalysed by the same cytochrome P450 in the Turkish subjects.

Omission of the deconjugation step in urine analysis and the unaltered outcome of CYP2D6 phenotyping with dextromethorphan.

The present study was aimed at determining whether the deconjugation step in chemical analysis could be omitted without altering the outcome of phenotyping CYP2D6 with dextromethorphan. This drug and its metabolite, dextrorphan, were assayed by high-performance liquid chromatography (HPLC) in urine. Urinary levels of dextromethorphan and dextrorphan with and without enzymatic (beta-glucuronidase) treatment of urine and the metabolic ratios for dextromethorphan were determined in 45 subjects. Although the enzymatic treatment did not alter the urinary concentration of dextromethorphan in both phenotypes, it increased the urinary concentration of dextrorphan in both poor and extensive metabolizers by 3.7- and 12.8-fold, respectively. A urinary unconjugated dextromethorphan/unconjugated dextrorphan metabolic ratio of 2.00 and a total dextromethorphan/total dextrorphan metabolic ratio of 0.30, respectively, identified three poor metabolizers. Enzymatic treatment decreased the urinary antimode value. Moreover, the urinary metabolic ratio based on unconjugated dextrorphan and dextromethorphan correlated well with that based on assay of total dextrorphan and dextromethorphan (rs = 0.9458, P < 0.001). The results show that urinary analysis of dextrorphan and dextromethorphan omitting the enzymatic deconjugation step is a fast, reliable and sensitive method and could be used for studying CYP2D6 type genetic polymorphism in man.

Penetration of ofloxacin and ciprofloxacin in aqueous humor after topical administration.

BACKGROUND AND OBJECTIVE: To compare the aqueous humor levels of 0.3% ofloxacin and 0.3% ciprofloxacin containing eyedrops in patients with healthy cornea. PATIENTS AND METHODS: Fifty patients with cataract were randomly assigned to have 0.3% ofloxacin containing eyedrop (25 patients) or 0.3% ciprofloxacin containing eyedrop (25 patients). Both drugs were repetitively instilled to each patient for 6 hours before the surgery. Aqueous samples were collected after penetrating the anterior chamber during cataract extraction and assayed by high-performance liquid chromatography method. RESULTS: The aqueous humor level of ofloxacin (1.43 +/- 0.26 microg/ml, mean +/- SEM) was significantly higher than that of ciprofloxacin (0.35 +/- 0.07 microg/ml) following the topical application (P < .0002). CONCLUSION: Aqueous humor penetration of topical ofloxacin is about 4 times higher than that of topical ciprofloxacin when the drugs are applied as described above.

S-mephenytoin, sparteine and debrisoquine oxidation: genetic polymorphisms in a Turkish population.

A mephenytoin test was carried out in 106 unrelated healthy Turkish volunteers. Racemic mephenytoin was coadministered with either debrisoquine or sparteine. The S/R mephenytoin ratio ranged from < 0.1 to 0.73 in 105 subjects, accordingly phenotyped as extensive metabolisers. One subject had an S/R mephenytoin ratio of 1.02, showing that he was a poor metaboliser of mephenytoin (0.94%, confidence interval 0.25% and 13.65%). In 48 subjects, the metabolic ratios of debrisoquine and sparteine were correlated significantly (rs = 0.61, P < 0.001).

Sulphamethoxazole acetylation in fast and slow acetylators.

Acetylation of sulphamethoxazole was studied in 22 subjects previously phenotyped with sulphadimidine. Sulphamethoxazole and its acetylated metabolite were measured in 6 h plasma and 0-6 h urine samples. Percentage of plasma acetylated-sulphamethoxazole did not correlate with the percentage of urinary acetylated-sulphamethoxazole. There was also no correlation between the percentage of acetylated-sulphadimidine and percentage of acetylated-sulphamethoxazole in plasma. The finding suggest that the acetylation of sulphamethoxazole has no predictive value in determining acetylator status.

N-acetylation phenotyping with sulphadimidine in a Turkish population.

The distribution of acetylator phenotypes was studied in 244 unrelated Turkish subjects. Sulphadimidine and its acetylated metabolite were measured in 6 h plasma and 0-6 h urine samples after an oral dose of 10 mg/kg. Subjects with 37.5% or less acetylsulphadimidine in plasma were regarded as slow acetylators and the others as rapid acetylators. The mean plasma concentration of acetylsulphadimidine was about 2.5-times lower in slow acetylators. Urinary excretion of total sulphadimidine (free + acetylated) was also significantly lower in slow acetylators compared to rapid acetylators. The frequency of slow acetylators was 60.7% in the population (95% confidence interval 54.3% to 66.8%). Sulphadimidine acetylation showed no variation due to sex, age, body weight or pre-existing disease.

Proguanil metabolism in relation to S-mephenytoin oxidation in a Turkish population.

The oxidation of proguanil was studied in 89 unrelated healthy Turkish volunteers after administration of proguanil (single dose, 200 mg, orally). Based on the distribution of the ratio of proguanil to cycloguanil excreted in urine, and using an antimode value of 15, the prevalence of poor metabolizers in a Turkish population was estimated to be 5.6% (95% confidence interval 2.0%-17.3%) which was similar to that in the other Caucasian populations. The relationship between the oxidative capacities of CYP2C19 for the two substrates, proguanil and mephenytoin, was studied in 39 subjects (two poor and 37 extensive metabolizers of proguanil). The two poor metabolizers of proguanil were also identified as poor metabolizers of S-mephenytoin and no misclassification by the two phenotyping methods was observed. The correlation between the metabolic ratio of proguanil to cycloguanil and the S/R-mephenytoin ratio as assessed by Spearman's rank test, was statistically significant (rs = 0.50, P < 0.001).

The hydroxylation of omeprazole correlates with S-mephenytoin and proguanil metabolism.

OBJECTIVES: This pharmacogenetic study was aimed at studying the pattern of oxidation of omeprazole in a Turkish population and testing whether omeprazole metabolism cosegregates with the genetically determined metabolism of mephenytoin and proguanil in Turkish subjects. METHODS: The hydroxylation of omeprazole was measured in 116 unrelated healthy Turkish subjects after administration of a single oral dose of omeprazole (20 mg), using the ratio of omeprazole to 5-hydroxyomeprazole in plasma 3 h after dosing. To 31 subjects, who were phenotyped with omeprazole, mephenytoin (100 mg, p.o.) or proguanil (200 mg, p.o.) were administered at least 1 week apart. The S/R ratio of mephenytoin and the ratio of proguanil to cycloguanil were determined from an 8-h urine collection. RESULTS: Based on the distribution of the log (omeprazole/hydroxyomeprazole) values and using the antimode value of 0.8, the frequency of poor metabolizers of omeprazole was estimated to be 7.7% (95% confidence interval 3-18%) which was similar to that in the other Caucasian populations (P = 0.54, Fisher's exact test). Three poor metabolizers of omeprazole were also classified as poor metabolizers of both mephenytoin and proguanil and no misclassification occurred with three phenotyping methods. All three methods separated poor or extensive metabolizer phenotypes with complete concordance. The ratio of omeprazole to hydroxyomeprazole correlated with the S/R ratio of mephenytoin and the ratio of proguanil to cycloguanil. CONCLUSION: These results support the hypothesis that the oxidative metabolism of three different drugs may be catalyzed by the same cytochrome P450 enzyme.

Penetration of ofloxacin in human aqueous and vitreous humors following oral and topical administration.

PURPOSE: To determine aqueous and vitreous humor ofloxacin levels following oral and topical application of ofloxacin in patients with noninflamed cornea and intact crystalline lens, and to compare the drug levels provided by each route. MATERIALS AND METHODS: Twenty-six patients undergoing pars plana vitrectomy for various ocular pathologies were divided into two groups. Fourteen patients received two drops of 0.3% ophthalmic solution of ofloxacin every 30 minutes for 3 hours and every 60 minutes for the next 3 hours, and 12 patients received a single oral dose of 400 mg ofloxacin 8 hours before surgery. The aqueous and vitreous humor samples were simultaneously collected after oral or topical administration during pars plana vitrectomy to assess penetration of the drug. Samples were assayed for ofloxacin concentrations by a previously described method using high-performance liquid chromatography. RESULTS: The aqueous and vitreous humor levels of ofloxacin were 1.54 +/- 0.27 microg/mL (mean +/- standard error) and 1.77 +/- 0.24 microg/mL after oral and 1.44 +/- 0.24 microg/mL and 0.37 +/- 0.05 microg/mL after topical ofloxacin administration, respectively. Aqueous humor levels were not statistically different following oral or topical administration (P > 0.8). However, vitreous level of the drug after oral administration was significantly higher than that after topical administration (P < 0.001). CONCLUSION: Ocular bioavailability of ofloxacin in aqueous humor after oral and topical administration is similar when the drug is applied as described. Penetration of ofloxacin into vitreous humor is less than that into aqueous humor following topical application. The aqueous humor levels of ofloxacin via both routes and the vitreous level of the drug after oral route exceed the minimum inhibitory concentrations for certain bacterial species that frequently cause intraocular infection.