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The underlying cause of tuberculosis (TB) treatment failure is still largely unknown. A 1 H NMR approach was applied to identify and quantify a subset of TB drugs and drug metabolites: ethambutol (EMB), acetyl isoniazid (AcINH), isonicotinic acid, pyrazinamide (PZA), pyrazinoic acid and 5-hydroxy-pyrazinoic acid, from the urine of TB patients. Samples were collected before, during (weeks one, two and four) and after standardised TB treatment. The median concentrations of the EMB and PZA metabolites were comparable between the samples from patients with eventually cured and failed treatment outcomes. The INH metabolites showed comparatively elevated concentrations in the treatment failure patients during and after treatment. Variation in INH metabolite concentrations couldn't be associated with the varying acetylator genotypes, and it is therefore suggested that treatment failure is influenced more so by other conditions, such as environmental factors, or individual variation in other INH metabolic pathways., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Alpibectir (also known as BVL-GSK098 and GSK3729098) is a new chemical entity (NCE) with a novel mechanism for the treatment of tuberculosis. The disposition of alpibectir was determined in subjects from a first-time-in-human trial after a single oral dose of 40 mg and after 7 days repeat dosing at 30 mg. Here we present a combined approach of 19 F-NMR (nuclear magnetic resonance), 1 H-NMR, and high-resolution mass spectrometry (HRMS) to confidently determine the human metabolic fate of alpibectir. Utilizing multiple sites of fluorination in the molecule, it was possible to fractionate human urine and plasma to confidently detect and quantify the metabolite responses using 19 F-NMR. Qualitative detection and structural characterization of F-containing NMR fractions were performed using complementary high-resolution ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) analyses to further add confidence to the metabolite responses in these fractions. Subsequent 1 H-NMR then provided unequivocal standard-free structural confirmation for key metabolites, which would not be possible with conventional radioactivity detection and LC-MS/MS techniques. Alpibectir was shown to undergo extensive hydrolysis of the central amide moiety, where the resultant N -dealkylated amine and trifluorobutyric acid products were detected initially by unbiased 19 F-NMR detection along with major downstream biotransformations to form a carbamoyl glucuronide conjugate and trifluoroacetic acid, respectively. Parallel UHPLC-MS/MS analyses provided confirmatory or additional structural characterization only where relevant. These concerted data allowed for the qualitative metabolic profile and quantitative determination of drug-related material (DRM) in urine and plasma, along with the percentage of dose excreted in urine, to be reported in a comprehensive, efficient, and data-led manner. SIGNIFICANCE STATEMENT: Combining the selectivity of 19 F-NMR (nuclear magnetic resonance) for unfractionated samples as first-intent, data-led sample fractionation prior to 19 F-NMR and structure-rich 1 H-NMR detection, along with the sensitivity of high-resolution ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS), a novel alternative for time-efficient detection and quantification of drug-related material (DRM) in human without use of radiolabeled drug is reported. This allowed more complete data rationalization of human metabolism, permitting early risk assessment and progression of the development of antitubercular agent, alpibectir., (Copyright © 2024 by The Author(s).)

Whole-body physiologically based pharmacokinetic (WB-PBPK) models have become an important tool in drug development, as they enable characterization of pharmacokinetic profiles across different organs based on physiological (systems-specific) and physicochemical (drug-specific) properties. However, it remains unclear which data are needed for accurate predictions when applying the approach to novel candidate molecules progressing into the clinic. In this work, as case study, we investigated the predictive performance of WB-PBPK models both for prospective and retrospective evaluation of the pharmacokinetics of ethambutol, considering scenarios that reflect different stages of development, including settings in which the data are limited to in vitro experiments, in vivo preclinical data, and when some clinical data are available. Overall, the accuracy of PBPK model-predicted systemic and tissue exposure was heavily dependant on prior knowledge about the eliminating organs. Whilst these findings may be specific to ethambutol, the challenges and potential limitations identified here may be relevant to a variety of drugs, raising questions about (1) the minimum requirements for prospective use of WB-PBPK models during the characterization of drug disposition and (2) implication of uncertainty for dose selection in humans., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Delamanid (OPC-67683, Deltyba™, nitro-dihydro-imidazooxazoles derivative) is approved for the treatment of adult pulmonary multidrug-resistant tuberculosis. The absorption, distribution and excretion of delamanid-derived radioactivity were investigated after a single oral administration of 14 C-delamanid at 3 mg/kg to rats. In both male and female rats, radioactivity in blood and all tissues reached peak levels by 8 or 24 h post-dose, and thereafter decreased slowly. Radioactivity levels were 3- to 5-fold higher in lung tissue at time to maximum concentration compared with plasma. In addition, radioactivity was broadly distributed in various tissues, including the central nervous system, eyeball, placenta and fetus, indicating that 14 C-delamanid permeated the brain, retinal and placental blood barriers. By 168 h post-dose, radioactivity in almost all the tissues was higher than that in the plasma. Radioactivity was also transferred into the milk of lactating rats. Approximately 6% and 92% of radioactivity was excreted in the urine and feces, respectively, indicating that the absorbed radioactivity was primarily excreted via the biliary route. No significant differences in the absorption, distribution and excretion of 14 C-delamanid were observed between male and female rats. The pharmacokinetic results suggested that delamanid was broadly distributed to the lungs and various tissues for a prolonged duration of time at concentrations expected to effectively target tuberculosis bacteria. These data indicate that delamanid, in addition to its previously demonstrated efficacy in pulmonary tuberculosis, might be an effective therapeutic approach to treating extrapulmonary tuberculosis. Copyright © 2017 John Wiley & Sons, Ltd., (Copyright © 2017 John Wiley & Sons, Ltd.)

Herein we describe the development, characterization and application of an electrochemical sensor based on the use of Nafion/MWCNT-modified screen-printed carbon electrodes (SPCEs) for the voltammetric detection of the anti-tuberculosis (anti-TB) drug ethambutol (ETB). The electrochemical behaviour of the drug at the surface of the developed Nafion/MWCNT-SPCEs was studied through cyclic voltammetry (CV) and square wave voltammetry (SWV) techniques. Electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM) were employed to characterize the modified surface of the electrodes. Results showed that, compared to both unmodified and MWCNTs-modified SPCEs, negatively charged Nafion/MWCNT-SPCEs remarkably enhanced the electrochemical sensitivity and selectivity for ETB due to the synergistic effect of the electrostatic interaction between cationic ETB molecules and negatively charged Nafion polymer and the inherent electrocatalytic properties of both MWCNTs and Nafion. Nafion/MWCNT-SPCEs provided excellent biocompatibility, good electrical conductivity, low electrochemical interferences and a high signal-to-noise ratio, providing excellent performance towards ETB quantification in microvolumes of human urine and human blood serum samples. The outcomes of this paper confirm that the Nafion/MWCNT-SPCE-based device could be a potential candidate for the development of a low-cost, yet reliable and efficient electrochemical portable sensor for the low-level detection of this antimycobacterial drug in biological samples.

Background: The cost and complexity of current approaches to therapeutic drug monitoring during tuberculosis (TB) therapy limits widespread use in areas of greatest need. We sought to determine whether urine colorimetry could have a novel application as a form of therapeutic drug monitoring during anti-TB therapy., Methods: Among healthy volunteers, we evaluated 3 dose sizes of rifampin (150 mg, 300 mg, and 600 mg), performed intensive pharmacokinetic sampling, and collected a timed urine void at 4 h post-dosing. The absorbance peak at 475 nm was measured after rifamycin extraction. The optimal cutoff was evaluated in a study of 39 HIV/TB patients undergoing TB treatment in Botswana., Results: In the derivation study, a urine colorimetric assay value of 4.0 × 10(-2) Abs, using a timed void 4 h after dosing, demonstrated a sensitivity of 92 % and specificity of 60 % to detect a peak rifampin concentration (Cmax) under 8 mg/L, with an area under the ROC curve of 0.92. In the validation study, this cutoff was specific (100 %) but insensitive (28 %). We observed similar test characteristics for a target Cmax target of 6.6 mg/L, and a target area under the drug concentration-versus-time curve (AUC0-8) target of 24.1 mg•hour/L., Conclusions: The urine colorimetric assay was specific but insensitive to detect low rifampin serum concentrations among HIV/TB patients. In future work we will attempt to optimize sampling times and assay performance, with the goal of delivering a method that can translate into a point-of-care assessment of rifampin exposure during anti-TB therapy.

Population level variation of drug metabolism phenotype (DMP) has great implications in treatment outcome, drug-related side effects, and resistance development. In this study, we used a gas chromatography-time of flight-mass spectrometry (GC-TOF-MS)-based untargeted urine metabolomics approach to understand the DMP of a tuberculosis (TB) patient cohort (n= 20) from Tripura, a state in the northeastern part of India. Urine samples collected at different postdose time points (2 h, 6 h, 12 h, 24 h, 36 h, and 48 h) from these newly diagnosed TB patients receiving first-line anti-TB drugs were analyzed, and we have successfully detected three of the four first-line drugs,viz, isoniazid (INH), ethambutol (ETB), and pyrazinamide (PZA). The majority of their known metabolites, acetyl-isoniazid (AcINH), isonicotinic acid (INA), isonicotinuric acid (INTA), 2,2'-(ethylenediimino)-dibutyric acid (EDBA), 5-hydroxypyrazinamide (5OH-PZA), pyrazinoic acid (POA), and 5-hydroxypyrazinoic acid (5OH-POA), were also detected. Analyzing the variation in abundances of drugs and their known metabolites and calculating the metabolic ratios in these samples, we offer comprehensive DMP information on this small patient cohort that represents Tripura, India. The majority (75%) of these patients are found to be slow acetylators of INH. The average metabolic ratios of POA/PZA and 5OH-POA/POA are 3.16 ± 3.03 and 6.09 ± 6.15, respectively. Employing correlation analysis of the metabolomics metadata and a manual prediction of drug catabolism, we have proposed 2-aminobutyric acid (AABA) as a novel metabolite of ETB. These observations indicate the usefulness of GC-MS-based metabolomics to characterize the DMP at a population level and also to identify novel drug metabolites., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

A novel colorimetric nanomaterial-assisted optical sensor for pazufloxacin mesilate was proposed for the first time. Pazufloxacin mesilate could induce the aggregation of glucose-reduced gold nanoparticles (AuNPs) through hydrogen-bonding interaction and electrostatic attraction, leading to the changes in color and absorption spectra of AuNPs. The effect of different factors such as pH, the amount of AuNPs, reaction time and reaction temperature was inspected. Under the optimum condition, UV-vis spectra showed that the absorption ratio (A670/A532) was linear with the concentration of pazufloxacin mesilate in the range from 9×10(-8) mol L(-1) to 7×10(-7) mol L(-1) with a linear coefficient of 0.9951. This method can be applied to detecting pazufloxacin mesilate with an ultralow detection limit of 7.92×10(-9) mol L(-1) without any complicated instruments. Through inspecting other analytes and ions, the anti-interference performance of AuNP detection system for pazufloxacin mesilate was excellent. For its high efficiency, rapid response rate as well as wide linear range, it had been successfully used to the analysis of pazufloxacin mesilate in human urine quantificationally., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The Arkansas colorimetric method monitors adherence to isoniazid (INH) by the detection of INH metabolites in urine. Urine samples 4 h after INH administration in 31 human immunodeficiency virus infected children receiving daily or thrice weekly INH preventive therapy were Arkansas test-positive for 29/31 (94%), while acetylisoniazid (AcINH) was detected in 30/31 (97%) using mass spectrometry. At 24, 48 and 72 h, only 78%, 23% and 0 samples, respectively, were Arkansas-positive, while INH or AcINH was detected in respectively 94%, 69% and 33%. The Arkansas test reliably predicted INH ingestion at a clinic visit 4 h after morning doses, but did not perform well at 24 h.

In this current study we used electrochemically active film which contains poly-L-methionine (PMET) and electrochemically reduced graphene oxide (ERGO) on glassy carbon electrode (GCE) for pyrazinamide (PZM) detection. The electrocatalytic response of analyte at PMET/ERGO/GCE film was measured using both cyclic voltammetry (CV) and differential pulse voltammetry (DPV). In addition, electrochemical impedance studies revealed that the smaller R(ct) value observed at PMET/ERGO film modified GCE which authenticates its good conductivity and faster electron transfer rate. The prepared PMET/ERGO/GCE film exhibits excellent DPV response towards PZM and the reduction peak current increased linearly with respect to PZM concentration in the linear range between 0.4 μM to 1129 μM with a sensitivity of 0.266 μA μM(-1) cm(-2). Real sample studies were carried out in human blood plasma and urine samples, which offered good recovery and revealed the promising practicality of the sensor for PZM detection. The proposed sensor displayed a good selectivity, repeatability, sensitivity with appreciable consistency and good reproducibility. In addition, the proposed electrochemical sensor showed good results towards the commercial pharmaceutical PZM samples., (Copyright © 2013 Elsevier Inc. All rights reserved.)

A simple and fast method for spectrophotometric determination of sparfloxacin using p-dimethyl-aminobenzaldehyde (DMAB) has been developed. A yellow coloured product formed from reaction between sparfloxacin and DMAB as a result of condensation reaction at room temperature. The maximum absorbance was found at 392 nm with molar absorptivity of 4.9 × 10(3) L mol(-1) cm(-1). All parameters for the reaction, as concentration of DMBA reagent, molarity of sulphuric acid, and reaction temperature were studied. Under the conditions studied, a linear relationship between absorbance of the condensation product and concentration of sparfloxacin in the range of 2.0-80.0 μg mL(-1) was found with good correlation coefficient (0.9997). The limits of detection (LOD) and quantification (LOQ) for the proposed method were found to be 0.22 and 0.75 μg mL(-1) respectively. The repeatability and accuracy (model) of the method was studied at three different concentrations of sparfloxacin and found with value of relative standard deviation less than 2.0%. The method was found selective for determination of sparfloxacin in the presence of commonly used excipients in dosage forms. The developed method was validated statistically and applied successfully to the analysis of the drug in pure form, pharmaceutical preparations, and spiked blood plasma and urine samples with good accuracy (real) and precision. The percentage recovery was found from 99.0-100.0% with relative standard deviation less than 1%. The results of the proposed method were compared statistically with the results of literature HPLC method.

Background: Poor adherence to isoniazid (INH) preventive therapy (IPT) is an impediment to effective control of latent tuberculosis (TB) infection. TB patients who smoke are at higher risk of latent TB infection, active disease, and TB mortality, and may have lower adherence to their TB medications. The objective of our study was to validate IsoScreen and SmokeScreen (GFC Diagnostics, UK), two point-of-care tests for monitoring INH intake and determining smoking status. The tests could be used together in the same individual to help identify patients with a high-risk profile and provide a tailored treatment plan that includes medication management, adherence interventions, and smoking cessation programs., Methodology/principal Findings: 200 adult outpatients attending the TB and/or the smoking cessation clinic were recruited at the Montreal Chest Institute. Sensitivity and specificity were measured for each test against the corresponding composite reference standard. Test reliability was measured using kappa statistic for intra-rater and inter-rater agreement. Univariate and multivariate logistic regression models were used to explore possible covariates that might be related to false-positive and false-negative test results. IsoScreen had a sensitivity of 93.2% (95% confidence interval [CI] 80.3, 98.2) and specificity of 98.7% (94.8, 99.8). IsoScreen had intra-rater agreement (kappa) of 0.75 (0.48, 0.94) and inter-rater agreement of 0.61 (0.27, 0.90). SmokeScreen had a sensitivity of 69.2% (56.4, 79.8), specificity of 81.6% (73.0, 88.0), intra-rater agreement of 0.77 (0.56, 0.94), and inter-rater agreement of 0.66 (0.42, 0.88). False-positive SmokeScreen tests were strongly associated with INH treatment., Conclusions: IsoScreen had high validity and reliability, whereas SmokeScreen had modest validity and reliability. SmokeScreen tests did not perform well in a population receiving INH due to the association between INH treatment and false-positive SmokeScreen test results. Development of the next generation SmokeScreen assay should account for this potential interference.

One of the most effective and widely used antituberculosis (anti-TB) drugs is isoniazid (INH), a prodrug activated via oxidation that forms an adduct with NAD(+) to inhibit NADH-dependent targets of Mycobacterium tuberculosis, such as enoyl-acyl carrier protein reductase (InhA). The metabolic by-products and potentially toxic intermediates resulting from INH therapy have been identified through a large body of work. However, an INH-NAD adduct or structures related to this adduct have not been identified in specimens from human TB patients or animal models of TB. Analyses by mass spectrometry of urine collected from TB patients in a study conducted by the NIAID-funded Tuberculosis Research Unit identified 4-isonicotinoylnicotinamide (C(12)H(9)N(3)O(2)) as a novel metabolite of INH therapy. This compound was formed by M. tuberculosis strains in a KatG-dependent manner but could also be produced by mice treated with INH independent of an M. tuberculosis infection. Thus, the 4-isonicotinoylnicotinamide observed in human urine samples is likely derived from the degradation of oxidized INH-NAD adducts and provides direct evidence of host INH activation.

Isoniazid (INH) is a first-line drug for tuberculosis control; the side effects of INH are thought to be associated with its metabolism, and this study was designed to globally characterize isoniazid metabolism. Metabolomic strategies were used to profile isoniazid metabolism in humans. Eight known and seven novel INH metabolites and hydrazones were identified in human urine. The novel products included two hydroxylated INH metabolites and five hydrazones. The two novel metabolites were determined as 2-oxo-1,2-dihydro-pyridine-4-carbohydrazide and isoniazid N-oxide. Five novel hydrazones were produced by condensation of isoniazid with keto acids that are intermediates in the metabolism of essential amino acids, namely, leucine and/or isoleucine, lysine, tyrosine, tryptophan, and phenylalanine. This study enhances our knowledge of isoniazid metabolism and disposition and may offer new avenues for investigating INH-induced toxicity.

Rifabutin, an anti-mycobacterial agent, is reported to be extensively metabolized in vivo into more than 20 biotransformation products, with similar profile both in humans and rats. Among the metabolites formed, only seven have been characterized, the remaining are unknown. Hence, the purpose of the present study was to fill this gap by using modern in silico tools combined with advanced liquid chromatography-mass spectrometry (LC-MS) techniques. Initially a comprehensive mass fragmentation pattern for rifabutin was established using Frontier 5.1 software coupled with the data collected from multiple-stage MS (MS(n)), high resolution MS (HR-MS) and hydrogen/deuterium exchange MS (HDE-MS) experiments. The metabolites were then predicted in silico by using different software like MetaSite, Metabolite Predict and MetWorks. The in silico results were verified through in vivo studies by administration of 20mg/kg drug to rats followed by LC-MS analyses. The urine was collected post-dose at different time intervals, and subjected to sample preparation involving sequentially protein-precipitation, liquid-freeze separation, and solid-phase extraction. The drug and metabolites were separated on an HPLC column followed by LC-MS studies. The difference of accurate masses of the drug and metabolites, and differences in their mass fragmentation pattern helped to assign structures to the metabolites and define the site of change. Here also, in silico detection tools were used, which provided complementary information. Using this strategy, 23 metabolites were detected and identified in rat urine without their isolation. The new sixteen metabolites were monohydroxy (05), dihydroxy (04), N-dealkyl (01), 25-O-desacetyl-27-O-demethyl (01), 25-O-desacetyl-23-O-acetyl (01), 25-O-desacetyl-monohydroxy (01), 27-O-demethyl-monohydroxy (01) and dehydrogenated (02) rifabutin., (Copyright (c) 2010 Elsevier B.V. All rights reserved.)

We evaluated the accuracy of a point-of-care test designed to measure adherence to isoniazid (INH) preventive therapy in a hospital setting in Rio de Janeiro, Brazil. Patients on treatment with daily INH and patients not receiving INH were included. Sensitivity and specificity of the test were 84%/98% at the first minute, and 95%/98% at the fifth minute, respectively. Among smokers, sensitivity and specificity was reduced (80%/89% at the fifth minute, respectively), but only 17% smoked. This test accurately detected INH metabolites 24h following directly observed INH intake, though sensitivity and specificity may be compromised by tobacco smoke exposure.

The Arkansas method (AM) for isoniazid (INH) metabolite detection is a relatively inexpensive, simple, objective measure of adherence. The purpose of the study was to explore whether variations in urine sample handling and storage will produce accurate assay outcomes. Participants were a convenience sample of 28 adults and adolescents prescribed INH for latent tuberculosis infection. Participants provided one sample to test effects of the following: mixing processes; durations at room temperature, in a refrigerator, or frozen; and effects of freeze/thaw cycles on AM outcomes. No manipulations had a discernible impact on outcomes with concordant positive rates from 85% to 100%. Concordance rates of manipulated samples did not appear to differ from rates of norm samples. Results suggest that urine samples can withstand a variety of manipulations in both handling and storage without affecting the accuracy of AM assay results. These findings have important implications for providers of treatment and researchers and provide the impetus for both to examine the potential of using the AM of INH metabolite testing as a measure of medication adherence.

A sensitive voltammetric method for trace measurements of isoniazid (INZ) in synthetic human urine sample is described. The method is based on its electroreduction at -0.98V vs. Ag/AgCl on screen-printed carbon electrode (SPCE) modified with poly-l-histidine (PH). A film of good adherence on SPCE and electrocatalytic properties was obtained coating the electrode by histidine monomer electropolymerization process (SPCE/EPH). The electrochemical behavior of the modified SPCE was investigated by cyclic, linear sweep (LSV), differential pulse (DPV), square-wave voltammetry (SWV), and electrochemical impedance spectroscopy (EIS). Limits of detection of 5.0x10(-7) mol L(-1), 1.7x10(-7) mol L(-1) and 2.5x10(-7) mol L(-1) were estimated from LSV, DPV and SWV determinations, respectively. The method was successfully applied to the determination of INZ in human urine samples., (2009 Elsevier B.V. All rights reserved.)

Setting: A South African hospital serving gold mine employees., Objective: To determine the sensitivity and specificity of the Arkansas method for detecting isoniazid (INH) metabolites among South African adults and to examine the effect of smoking status on positive results., Design: Urine specimens were collected from in-patients taking INH as part of tuberculosis treatment at 6, 12 and 24 h after a directly observed 300 mg oral dose. As a control group, a single urine specimen was collected from surgical in-patients not taking INH. Specimens were tested for INH using a commercially available dipstick., Results: A total of 153 patients on INH and 60 controls were recruited. The sensitivity of the test was 93.3% (95%CI 88.1-96.8) at 6 h post INH, 93.4% (95%CI 88.2-96.8) at 12 h and 77% (95%CI 69.1-83.7) at 24 h. The specificity of the test was 98.3% (95%CI 91.1->99.9). There was no association between smoking status and colour change of positive results., Conclusions: This test is a useful method of monitoring adherence to TB treatment or preventive therapy among South Africans. However, it is less than 100% sensitive, especially with increasing time post dose, which should be taken into consideration when interpreting results for individual patients.

Background & Objective: AIDS and its associated gastrointestinal complications may impair the absorption of anti-tuberculosis (TB) drugs. Impaired absorption of anti-TB drugs could lead to low drug exposure, which might contribute to acquired drug resistance and reduced effectiveness of anti-TB treatment. The aim of this study was to obtain information on the status of absorption of rifampicin (RMP) and isoniazid (INH) in asymptomatic HIV- positive individuals, who are less immunocompromised. The D-xylose absorption test was also carried out to assess the absorptive capacity of intestive., Methods: The absorption of RMP, INH and D-xylose was studied in 15 asymptomatic HIV-positive individuals with CD4 cell counts>350 cells/mm3 and 16 healthy volunteers, after oral administration of single doses of RMP (450 mg), INH (300 mg) and D-xylose (5 g). Urine was collected up to 8 h after drug administration. Percentage dose of the drugs and their metabolites and D-xylose excreted in urine were calculated., Results: A significant reduction in the urinary excretion of INH and D-xylose in HIV-positive persons compared to healthy volunteers was observed. The per cent dose of RMP and its metabolite, desacetyl RMP was also lower in HIV-positive persons compared to healthy volunteers, but this difference was not statistically significant., Interpretation & Conclusion: Decreased urinary excretion of D-xylose and INH are suggestive of intestinal malabsorption in HIV-positive individuals. HIV infection could cause malabsorption of anti-TB drugs even at an early stage of the disease. The clinical implications of these findings need to be confirmed in larger studies.

Background: Tuberculosis (TB) is the most common opportunistic infection in HIV-infected adults in developing countries. Isoniazid (INH) is recommended for treatment of latent TB infection, however non-adherence is common. The purpose of this study was to apply in-house prepared isoniazid (INH) urine test strips in a clinical setting, and identify predictors of positive test results in an adherence questionnaire in HIV-infected adults taking INH for prevention of TB., Methods: Cross-sectional study of adherence using a questionnaire and urine test strips for detection of INH metabolites at two hospitals in Pietermaritzburg, South Africa. Participants were aged at least 18 years, HIV positive, and receiving INH for prevention of tuberculosis disease. Univariate and multivariate analyses are used to identify factors relevant to adherence., Results: 301 consecutive patients were recruited. 28% of participants had negative urine tests. 32 (37.2%, 95% CI25.4, 45.0) of the 86 patients who received INH from peripheral pharmacies said the pharmacy had run out of INH at some time, compared with central hospital pharmacies (p = 0.0001). In univariate analysis, a negative test was associated with self-reported missed INH doses (p = 0.043). Each 12-hour increment since last reported dose increased the likelihood of a negative test by 34% (p = 0.0007). Belief in INH safety was associated with a positive test (p = 0.021). In multivariate analysis, patients who believed INH is important for prevention of TB disease were more likely to be negative (p = 0.0086)., Conclusion: Adequate drug availability at peripheral pharmacies remains an important intervention for TB prevention. Key questions may identify potentially non-adherent patients. In-house prepared urine tests strips are an effective and cheap method of objectively assessing INH adherence, and could be used an important tool in TB control programs.

This investigation retrospectively assessed inexpensive non-invasive qualitative methods to monitor the ingestion of anti-tuberculosis drugs isoniazid, rifampicin and rifapentine. Results showed that commercial test strips detected the isoniazid metabolites isonicotinic acid and isonicotinylglycine as efficiently as the isonicotinic acid method in 150 urine samples. The presence of rifamycins in urine samples (n=1085) was detected by microbiological assay techniques and the sensitivity compared to the n-butanol extraction colour test in 91 of these specimens. The proportions detected by the two methods were significantly different and the sensitivity of the n-butanol procedure was only 63.8% (95% CL 51.2-76.4%) as compared to that of the superior microbiological method. Final validation (n=691) showed that qualitative assays measure isoniazid and rifamycin ingestion with an efficiency similar to high-performance liquid chromatography. The qualitative procedures may therefore be valuable in clinical trials and in tuberculosis clinics to confirm drug ingestion.

Background: Adherence with therapy is a wide spectrum of behavior rather than a categorical state. While extreme nonadherence is readily apparent, it is rare compared to lesser degrees of nonadherence, which are difficult to predict., Aims: To compare the accuracy of doctor, nurse, and patient prediction of adherence with antituberculous therapy with urine isoniazid levels., Methods: A prospective, blinded clinical study was conducted, comparing adherence to antituberculous therapy as reported by patients, doctors, and nurses with urine isoniazid levels. We studied 173 patients with active tuberculosis (TB) recruited over 3 years in two TB clinics in Victoria, Australia. Adherence was defined as six random urine isoniazid (INH) levels being >0. Blinded assessment of adherence was completed by doctors, nurses, and patients. Lid opening and closing of computerized pill bottles were measured in a random subsample., Results: Of 173 patients, the rate of nonadherence was 24% (41/173) by urine INH, 54% (93/173) by patient self-report, 11% (19/173) by doctor assessment, and 7% (12/173) by nurse assessment. The sensitivity of prediction of nonadherence was 76% for patient self-report, 24% for doctor assessment, and 19% for nurse assessment. The 10 patients who used computerized pill bottles were all (100%) noncompliant at some stage., Conclusion: Nonadherence is common and poorly predicted by doctors and nurses, even those with extensive experience in treating TB. Contrary to popular belief, patient self-report is more reliable than doctor or nurse assessment of nonadherence. As clinicians, asking patients about adherence may be more valuable than attempting to judge for ourselves.

Background: In the fight against the global tuberculosis epidemic, it is essential to ensure that patients adhere to the treatment prescribed. As the treatment is given for a minimum of 6 months it is common for patients not to take their drugs regularly. Strategies are therefore needed to assess adherence to treatment. One established method is to examine the patient's urine for the presence of drug metabolites. A rapid point-of-care test would overcome some of the drawbacks associated with currently available methods., Method: A rapid, safe point-of-care test for isoniazid metabolites (IsoScreen, Surescreen Diagnostics Limited, Derby, UK) has been developed and used to help assess adherence to treatment in a busy clinic for tuberculosis patients in South London., Results: Urine samples were examined from 191 patients receiving isoniazid, usually in combination with rifampicin and other anti-tuberculous drugs. Isoscreen was positive in 93.2% of patients, suggesting that 6.8% might be poorly adhering to treatment. By contrast, examining the same urine samples for evidence of rifampicin ingestion gave positive results in only 43.5%, due to the fact that this test is only positive for a few hours after drug ingestion., Conclusion: IsoScreen has been shown to provide a rapid and safe point-of-care test, which contributes to the detection of non-adherence in patients with tuberculosis.

Although accurate data are scarce for children, tuberculosis (TB) represents one of the most common infectious causes of morbidity and mortality worldwide. TB case rates have declined among children in the United States in the last decade, but they remain high among children from low-income countries and racial or ethnic minorities. Establishing the definitive diagnosis of TB in a child remains difficult and frequently relies on a constellation of history, clinical findings, and bacteriology. Recently, updated national and international treatment recommendations have been published. Contact investigation and treatment using directly observed therapy are important components of the optimal case detection and management of TB in children.

A simple column chromatographic method for determination of ethambutol (EMB) in pharmaceutical preparations containing EMB in combination with other anti-TB drugs is presented. The method involved extraction of EMB into an organic solvent, followed by basification and column chromatographic separation on Amberlite CG 50 (100-200 mesh) and elution with suitable eluants and estimation at a wavelength of 270 nm. The assay was linear from 25 to 400 microg/ml. The relative standard deviations of intra and inter day assays were lower than 5%. Ethambutol was recovered from human urine quantitatively and stable for a period of at least one week in urine stored at -20 degrees C.

Isoniazid, pyridine-4-carboxylic acid hydrazide, is an antituberculosis-agent, which is used to prevent the development of clinical tuberculosis. A validated square-wave adsorptive cathodic stripping voltammetric procedure for the trace determination of the bulk drug at the hanging mercury drop electrode (HMDE) has been developed. Under the optimized conditions, (accumulation potential=-0.9 V, accumulation time=50-300 s, scan increment=8 mV, pulse-amplitude=25 mV, frequency=120 Hz and acetate buffer at pH 5.5) isoniazed generated two irreversible cathodic peaks. The first peak current showed a linear dependence with the drug concentration over the range 5 x 10(-10)-21 x 0(-6) M. The mean percentage recoveries, based on the average of five replicate measurements, for 7 x 10(-9) and 5 x 10(-8) M isoniazid were 97.71+/-2.93 and 99.76+/-0.77, respectively. The achieved limits of detection (LOD) and quantitation (LOQ) were 1.18 x 10(-10) and 3.93 x 10(-10) M isoniazid, respectively. The procedure was applied to the assay of the drug in tablets (Isocid and T.B. Zide), spiked human serum and urine with mean percentage recoveries of 97.81+/-1.49, 97.45+/-2.09, and 97.08+/-1.06, respectively. The limits of detection of 1.47 x 10(-9) and 2.4 x 10(-8) M, and quantitation of 4.9 x 10(-9) and 8 x 10(-8) M drug in human serum and urine, respectively, were achieved. The mean values of the various pharmackinetic parameters of isoniazid (C(max), T(max), t(1/2), AUC, and K(e)), estimated from analysis of plasma of two volunteers by means of the proposed procedure were similar to literature values.

Objectives: We sought to determine the efficacy of coaching Latino adolescents with latent tuberculosis infection to adhere to isoniazid treatment., Methods: Participants (n = 286) were randomly assigned to adherence coaching, attention control, or usual care groups. Adherence was measured via interviews and validated with urine assays., Results: Coaching resulted in significant increases in adherence compared with attention and usual care groups. Bicultural adolescents were more likely to be adherent than those most or least acculturated. Age and risk behavior were negatively related to adherence., Conclusions: Coaching can increase Latino adolescents' adherence to treatment for latent tuberculosis infection and should contribute to tuberculosis control for adolescents at high risk of contracting the disease.

Setting: A randomised, controlled clinical trial of the effectiveness of a family-based programme of directly observed treatment (DOT) for tuberculosis., Methods: TB patients seen in Victoria, Australia, were randomly allocated to DOT observed by a family member (FDOT), or to standard supervised but non-observed therapy (ST). The outcome measure was compliance, measured by blinded testing of isoniazid levels in urine. An intention-to-treat analysis was used., Results: Of 173 patients, 87 were allocated to FDOT and 86 to ST. Only 58% in the FDOT group were able to receive FDOT, the major reason being living alone and not having a family member to observe treatment. The rate of non-compliance was 24% (41/173), with no significant difference between FDOT (22/87) and ST (19/86). No clinical or socio-demographic variable predicted compliance., Conclusions: We were unable to demonstrate a benefit of FDOT in an urban, industrialised country setting. FDOT may be more appropriate in developing countries, where extended family support is often available and the burden of TB is much higher. Poor compliance and the difficulty in predicting non-compliance shown in this study highlights the need for DOT for all TB patients.

Study Objective: To apply the Arkansas color method in order to evaluate drug compliance and factors that can predict treatment adherence in patients being treated for latent tuberculosis infection (LTBI) with a single daily dose of isoniazid (INH)., Design: Prospective study of adherence of 105 patients aged 1 to 75 years who were treated with a single daily dose of INH for LTBI., Interventions: Patients or their parents were interviewed regarding parameters that may affect compliance. Urine samples were collected and tested for INH metabolites with the Arkansas color method., Results: Nonadherence to treatment was found in 28.5% of patients. There was no statistically significant correlation among the following parameters: gender; age; diagnosis; mode of administration (self or parents); duration of treatment; dose of INH per weight; or interval since last intake of dose. Twenty-six patients were randomly checked for treatment adherence on two separate visits, and nonadherent patients were informed immediately and their condition was fully explained to them. Five of six patients who were nonadherent in the first visit and were examined twice became adherent in the second visit. Three of 20 patients who were adherent in the first visit became nonadherent., Conclusion: Almost one third of the patients who received LTBI treatment with INH were nonadherent to treatment. No factor was found to predict adherence. The Arkansas method can be used by the family physician and is a simple, immediate method to follow-up patients with LTBI who are treated with INH.

Patient compliance should be ensured in an effective tuberculosis control programme. We measured patient compliance by detecting antituberculous drugs in the urine of 237 outpatients receiving one to three antituberculous drugs. Positive controls were 20 hospitalised patients, supervised to receive isoniazid (INH), rifampicin (RIF) and pyrazinamide (PZA), and negative controls were not on any drugs. Among the 237 study patients, only 67% were found to be taking the appropriate treatment and 8% had taken none. We conclude that a remarkable number of patients (33%) were non-compliant with treatment. The detection of antituberculous drugs in the urine is a quick, simple and inexpensive means of measuring adherence to treatment. Unless directly observed therapy (DOT) is adopted, we recommend routine urine testing for antituberculous drugs to identify defaulting patients.

Setting: Mulago Hospital, Kampala, Uganda., Objective: To evaluate the usefulness of urine dipsticks for monitoring adherence to anti-tuberculosis chemotherapy., Design: In-house urine dipsticks for detection of isoniazid (INH) metabolites were compared to commercial test strips. The value of n-butanol to detect rifampicin was compared to coloration of the urine. Non-adherence was assessed through a questionnaire and reviews of the Mulago Hospital TB register., Results: Urine was obtained from 236 patients (127 adults and 109 children) on daily chemotherapy. Using commercial test strips as standard, the sensitivity of in-house urine dipsticks was 99.5% and specificity was 96.4%. The sensitivity and the specificity of n-butanol and of coloration of urine to detect rifampicin were low (64.0% and 54.9%, and 85.5% and 64.8%, respectively). Fifty patients (21.2%) admitted non-adherence to treatment during the previous month. An additional 15 (6.8%) were detected through urine testing. Of 911 patients in the TB register of Mulago Hospital who had started treatment in the first 3 months of 2000, 39.7% did not complete their treatment. Two-thirds of these had discontinued treatment in the first 2 months., Conclusion: In-house INH test strips are as effective as commercially available strips for detecting isoniazid in the urine. They are a simple tool for monitoring adherence. Adherence to anti-tuberculosis chemotherapy as determined by the use of isoniazid test strips and review of the TB register showed poor compliance. Tests for rifampicin are less sensitive and specific.

For the purpose of more accurate and rapid analysis of isoniazid in body fluid samples, a reversed-phase high performance liquid chromatographic (HPLC) method was developed. Vanillin, being as a derivatizing reagent, was added to the plasma and urine samples when they were pretreated. Isoniazid and vanillin reacted to form isonicotinoyl hydrazone which was separated and detected. The pretreatment method of sample, the linear range, the precision, and the recovery of isoniazid were investigated by using human's plasma and urine spiked with standard isoniazid. The linear range was 0.2 mg/L- 12.0 mg/L (for plasma, r =0.999 6; for urine, r = 0.999 4). The detection limit was 0.2 mg/L. The intra-day and inter-day precisions of assay for isoniazid were 2.3% - 2.6% and 3.8% - 4.0% (n = 5) for plasma, 1.3% - 4.0% and 2.1% - 3.9% (n = 5) for urine respectively. The average recoveries of isoniazid were from 96.5% to 99.8% in plasma and from 96.3% to 99.4% in urine. The HPLC method has been used to investigate the concentration of isoniazid in the volunteer's plasma. The quantitative analysis of isoniazid in plasma was not interfered by impurities, the metabolites of isoniazid and the derivatizing reagent residue. This analytical method for isoniazid is sensitive, rapid and convenient. It is suitable for the analysis of toxics in forensic samples, the detection of drug concentration in clinical medicines and pharmacokinetic studies.

This study examined the validity of the Arkansas urine test. One hundred ninety-four adolescents submitted an unannounced urine specimen monthly (for 6 to 8 months). Duplicate specimens were blindly tested with high agreement (kappa >90%). Sensitivity and specificity were estimated. In 68% of test runs, adolescents recalled taking INH within 24 hr of specimen collection. For recall intervals of 24, 48, and 72 hr, sensitivity was 87, 85, and 83%, respectively. Females were less likely to test positive when INH was taken within the previous 24 hr (sensitivity 84 versus 92% males). Specificity was 57, 91, and 95% at 24, 48, and 72 hr, respectively. The Arkansas urine test was practical to use, and results correlated well with self-reported adherence to INH for treatment of latent tuberculosis infection (LTBI), over several months of follow-up. The test may be useful as part of an adherence-monitoring program when used in conjunction with self-reported measures.

The objective of this study was to develop a new simplified method using thin-layer chromatography (TLC) for determining isoniazid (INH) compliance in patients receiving antituberculosis chemotherapy. TLC was performed on silica gel plates using a standard solution of INH and acetylisoniazid (AcINH) and ethyl acetate-methanol (70:30 v/v) as the developing solvent. The spots of compound were detected by iodine. In the human study, fractional urine samples were collected over 24 hours from 4 healthy human subjects genotyped for NAT2* and to whom 400 mg of INH were administered orally. These samples were used for TLC analysis. The results of TLC were compared with those of high-performance liquid chromatography (HPLC). This method indicated good separation between INH and AcINH in standard solutions. The detection limits for INH and AcINH (applied volume; 20 microl of standard solution) were 2.2 nmole and 5 nmole, respectively, as detected by iodine. In the human study, the INH spot in urine was not detected on the TLC plate, except in one sample over the 0- to 4-hour period from 1 volunteer. However, the AcINH spot was detected in all urine samples from all volunteers. The total experimental time from application of the urine sample to analysis on TLC was 30 minutes. The results suggest that this method for detecting AcINH on TLC is an excellent, convenient, and simple method for determining INH compliance in patients receiving standard antituberculosis chemotherapy regimen or INH preventative therapy, regardless of the patient's NAT2* genotype.

Salivary and urinary excretion and plasma-saliva concentration ratios of isoniazid (INH) in the absence and presence of ciprofloxacin (CP) were investigated in healthy female volunteers. Results obtained indicated an absorption form of interaction between INH and CP. This led to delay in gastric emptying and onset of absorption of INH in the upper part of the gastrointestinal tract, resulting in a corresponding delay in the onset of salivary and urinary excretion of the drugs. There was a 1-hour reduction in the time to attain peak saliva concentration of INH (tmax), an insignificant difference in peak saliva concentration (Cmax), and a significant (P = 0.05) increase in AUC(0-24h) of INH in the presence of CP. Cumulative amount of INH excreted in the urine increased approximately 38% in the presence of CP. The calculated plasma-saliva concentration ratios of INH were reduced in the presence of CP and were slightly lower than the experimental values. This indicates increased amount of the drug secreted into saliva in the presence of CP and possible buccal partitioning of the drug. Overall, results of the current study indicate that CP delayed the onset but not the extent of INH absorption. Therefore, concurrent administration of the two drugs was considered relatively safe, and the absorption interaction that may have occurred may not be of reasonable clinical consequence.

Setting: In sub-Saharan Africa, tuberculosis (TB) has increased over the last two decades due to the human immunodeficiency virus pandemic. In Malawi, 20630 new TB patients were notified to the National Tuberculosis Programme in 1996, a fourfold increase since 1986. Due to this increase in cases and lack of resources (both human and monetary) it is becoming more difficult to ensure directly observed treatment (DOT) in the TB wards., Methods: In Ntcheu district, Malawi, a new TB regimen was introduced from April 1996 in which patients received supervised treatment by either a health worker or a guardian (i.e., family member). Adherence to the different treatment options was measured by form checks, tablet counts, and tests for detecting isoniazid in the urine. Adherence was measured at 2, 4 and 8 weeks after onset of TB treatment., Results: Overall adherence rate was 95-96%. Inpatients showed the highest adherence rates. Patients on guardian-based DOT (GB-DOT) (n = 35) showed 94% adherence, while patients on health centre based DOT (n = 40) showed more non-adherent behaviour: 11% according to monitoring forms, 14% according to tablet counts and 16% according to urine tests., Discussion: The results suggest that decentralised care is a feasible option for anti-tuberculosis treatment and that guardians can supervise TB treatment just as well as health workers during the intensive phase of TB treatment.

Data regarding possible differences in microbiological response to therapy of disease caused by Mycobacterium tuberculosis and M. africanum are limited. Presenting clinical characteristics and sputum bacillary load during standard short-course chemotherapy in patients with newly-diagnosed pulmonary tuberculosis due to M. tuberculosis (n = 7) and M. africanum (n = 6) were compared. Changes in sputum bacillary load were measured using quantitative acid-fast bacilli smears, colony forming unit assay, and time until positive culture in the BACTEC radiometric system. Presentation and response to short course chemotherapy were comparable between patients infected with M. tuberculosis and those infected with M. africanum.

An improved high-voltage electric field isolating joint and small detection cell have been carefully designed and fabricated. The joint possesses short steady time, high electric conductance efficiency and high performance. The cell is convenient to install and remove the capillaries with and without the joint, as well as to fix, adjust and insert the microelectrode into the detection capillary. Using the joint and the cell, an analytical method for determination of ofloxacin (Oflx) and pasiniazid (Ipa) in urine by capillary electrophoresis with on-column amperometric detection was developed. The calibration lines were linear in the range of 10-100 mg l-1 of Oflx and 1.0-50 mg l-1 of Ipa, respectively. The detection limits were 8.5 mg l-1 of Oflx and 0.80 mg l-1 of Ipa. Their recovery ranged from 101 to 104%. The accuracy and intra-day and inter-day reproducibility of Oflx and Ipa were determined with satisfactory results. This method was successfully used for determining Oflx and Ipa in human urine.

The effects of pefloxacin (PFC), a fluoroquinolone antibiotic, on the urinary and salivary concentrations of Isoniazid (INH) were investigated in six healthy female volunteers 19 to 30 years of age. The presence of PFC increased the rate and extent of INH absorption and the rate of its excretion in the urine and saliva. There was an increase in the excretion rate constant (K) and a reduction in the half-life (t1/2) of INH in the presence of PFC. Four of the volunteers had t(1/2) values in the range of 1.55 to 2.43 hours and were considered to be fast acetylators, whereas two subjects with a t(1/2) in the range of 3.36 to 4.41 hours were considered to be slow acetylators. Concurrent administration of INH and PFC may lead to an increased INH toxicity based on the results of the present study.

Setting: The benefits of fixed-dose combination (FDC) formulations of rifampicin, isoniazid and pyrazinamide over individual formulations are well recognised., Objectives: To evaluate the comparative bioavailability of antituberculosis drugs in FDC formulations and the same doses in separate formulations of antituberculosis drugs, using a simplified protocol developed by the World Health Organization (WHO)., Methods: Twenty healthy volunteers were included in the study and evaluated for bioequivalence of rifampicin in a cross-over experimental design. After administration of drugs the plasma concentration of rifampicin and desacetyl-rifampicin was measured repeatedly up to 8 hours in both plasma and urine. Various pharmacokinetic parameters of rifampicin, such as Cmax, Tmax, elimination rate constant, area under the curve (AUC) up to 8 hours and absorption efficiency were calculated., Results: No significant differences were observed between the FDCs and separate formulations when Cmax, Tmax, AUC and absorption efficiencies were compared by parametric test and Hauschke's analysis., Conclusion: The WHO simplified protocol is suitable for evaluating bioequivalence of antituberculosis drugs.

The effect of repeated administration of rifabutin on the pharmacokinetics and metabolism of ethambutol was evaluated in ten healthy volunteers. The subjects received a single oral administration of 1200 mg ethambutol on days 1 and 10 and a single daily oral dose of 300 mg rifabutin from days 3 to 9. No statistically significant difference was found in plasma pharmacokinetics (C(max), t(max), AUC, half-life and MRT) and in the renal clearance, whereas a significant decrease in the amount of unchanged ethambutol excreted in urine was observed. The decrease observed in ethambutol urinary excretion may be accounted for by taking into consideration the variability of the urinary excretion of ethambutol reported in the literature. However, a slight, likely not clinically relevant, induction or activation of kidney alcohol and/or aldehyde dehydrogenase isoenzymes by rifabutin cannot be ruled out at present. Evidence exists in the present study for autoinduction of rifabutin metabolism; this is shown by the lower plasma concentrations obtained 24 h after the seventh dose as compared to the theoretical concentrations., (Copyright 1999 Academic Press.)

A simple, specific, sensitive, and rapid method has been developed and validated for the determination of sparfloxacin in human plasma and urine. The assay consisted of reversed-phase HPLC with ultraviolet detection. Plasma proteins were efficiently removed by precipitation with perchloric acid after the addition of grepafloxacin as an internal standard. For the urine samples, the only required sample preparation was dilution. Separation was achieved on a C18 reversed-phase column. The quantification limit was 0.025 mg/L in plasma and 0.5 mg/L in urine. The coefficients of variation (CV) were less than 10% for intra-day and inter-day analyses. The recovery of sparfloxacin added to plasma and urine ranged from 96.7% to 97.9%. The method has been successfully applied to pharmacokinetic studies.

Urinary tract infection (UTI) is a common illness, with > or =30% of all women experiencing a UTI during their lifetime. Less than a decade ago, the standard therapy for acute uncomplicated UTIs involved treatment with > or =7 days of an antibacterial agent, but recent studies using a variety of newly introduced antibiotics, including the fluoroquinolones, have demonstrated that a 1- to 5-day treatment regimen can be equally effective. This randomized, double-masked, multicenter study was conducted to compare the efficacy and tolerability of a single dose of sparfloxacin with those of a 3-day regimen of sparfloxacin and a 7-day regimen of ciprofloxacin in the treatment of women with community-acquired acute uncomplicated urinary tract infection. A total of 1175 women were enrolled; 395 received sparfloxacin as a single 400-mg dose on day 1, 394 received sparfloxacin as a 400-mg loading dose on day 1 followed by 200 mg once daily for 2 additional days, and 386 received ciprofloxacin 250 mg twice daily for 7 days. Patients were comparable with respect to demographic characteristics and underlying conditions. A total of 954 patients were clinically assessable; 490 of these were also bacteriologically assessable. All patients treated were included in the tolerability analysis. Escherichia coli (75.4%), Klebsiella pneumoniae (4.9%), Enterococcus faecalis (4.6%), and Staphylococcus saprophyticus (4.1%) were the most commonly isolated organisms. In the all-treated population, clinical success was achieved 5 to 9 days after therapy in 91.8%, 92.2%, and 91.6% of patients in the single-dose sparfloxacin, 3-day sparfloxacin, and 7-day ciprofloxacin groups, respectively; bacteriologic success was observed in 91.7%, 92.6%, and 96.6% of those in the 3 groups. Sustained clinical success rates 4 to 6 weeks after therapy were 76.6%, 80.2%, and 79.5% in the single-dose sparfloxacin, 3-day sparfloxacin, and 7-day ciprofloxacin groups, respectively; sustained bacteriologic success rates were 80.7%, 90.1%, and 92.6%. The most common adverse events were nausea, headache, vaginal thrush, dizziness, and diarrhea; >92% of adverse events were mild or moderate in severity. The 2 drugs had comparable frequencies of adverse events, except for photosensitivity, which occurred in 3.3% of the 3-day sparfloxacin group, 1.3% of the single-dose sparfloxacin group, and 0.3% of the ciprofloxacin group (P = 0.005). The 3-day sparfloxacin regimen was effective and well tolerated. The initial response to single-dose sparfloxacin treatment was comparable to the response to the other 2 regimens, but the single-dose regimen proved less effective over time, with higher rates of clinical recurrence and bacteriologic relapse. Sparfloxacin provides an alternative to ciprofloxacin for patients with acute uncomplicated urinary tract infection who are not at risk for photosensitivity reactions or adverse events associated with a prolonged corrected QT interval.

Setting: The present study assesses bioavailability indices for rifampicin, isoniazid and pyrazinamide when administered to healthy volunteers separately or in a fixed triple-drug formulation, Rifater 125 SCT., Objective: To compare the pharmacokinetics of rifampicin, isoniazid and pyrazinamide based on their blood concentrations up to 12 hours with the proportions of the doses of the drugs and their metabolites excreted in urine up to 12 hours, and to assess the bioavailability indices for the free and fixed triple drug formulations., Design: An open cross-over study was conducted in 18 healthy volunteers with normal hepatic and renal functions to whom the drug combinations were administered in free and fixed dose formulations a week apart, to the same subject., Results: Concentrations of the three drugs/metabolites were assessed in blood and urine. The results indicated the absence of negative pharmacokinetic interactions between the drugs when administered in both the free and the new fixed triple drug formulation., Conclusion: Human bioavailability studies provide direct straightforward information, particularly when studying compounds such as rifampicin and other major anti-tuberculosis drugs. The results of the present study indicate that the pharmacokinetic properties of rifampicin, isoniazid and pyrazinamide as assessed after individual and combined administration do not change when combined in a single pharmaceutical preparation. The bioavailability indices calculated based on plasma concentrations and urinary levels for all three drugs compared well.

Objective: To investigate the effect of acidification and alkalinization on the pharmacokinetics of sparfloxacin in healthy subjects., Methods: A single 200-mg oral dose of sparfloxacin was given to nine healthy Japanese volunteers on three separate occasions under different conditions of urinary pH. Acidic and alkaline conditions were achieved by repeated oral doses of ammonium chloride and sodium bicarbonate, respectively. The concentrations of sparfloxacin and its metabolite in plasma and urine were determined by high-performance liquid chromatography assays., Results: The difference between treatments for Cmax, AUCinfinity, and CL x f(-1) were found to be significant. The relative bioavailability of sparfloxacin was 84.4% and 122.3% after ammonium chloride and sodium bicarbonate treatments, respectively. The amount of unchanged sparfloxacin in urine samples collected 0-48 h after sparfloxacin administration represented 10.1% of the dose in the control, 14.3% of the dose in urine acidification and 8.4% of the dose with alkalinization of urine. Renal clearance was found to depend on urinary pH. However, the plasma elimination and the metabolism of sparfloxacin were not significantly altered by acidification or alkalinization of the urine., Conclusion: The urinary pH dependence of the renal clearance of sparfloxacin will be of minor clinical importance with regard to the low contribution of renal excretion to the overall elimination of sparfloxacin. On the other hand, the alteration in the environmental pH in the gastrointestinal tract, produced by the concomitant ingestion of ammonium chloride or sodium bicarbonate, influences the absorption and bioavailability of sparfloxacin. This effect is likely to be clinically significant.