Development of UV/HPLC Methods for Quantitative Analysis of Benznidazole in Human Plasma and Urine for Application in Pediatric Clinical Studies

Marson, María Elena; Dana, Diego Dante; Altcheh, Jaime; García-Bournissen, Facundo; Mastrantonio, Guido

doi:10.1002/jcla.21615

Cited by 16 publications

(19 citation statements)

References 18 publications

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“…The detection of this substance, although not identified, was previously reported by our work group (Marson et al 2013).…”

Section: Introductionsupporting

confidence: 56%

“…Plasma samples of all patients were obtained from venous blood and BNZ was measured, most of the patients, in three different moments of treatment: at starting, in steady state and after finishing treatment, with previous reported HPLC/UV method (Marson et al 2013). In these measures, the samples of patients in steady state showed a particular HPLC chromatographic pattern, compared with samples of patients at the beginning of the treatment.…”

Section: Patient Plasma Samples and Pretreatmentmentioning

confidence: 99%

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Identification of N-benzylacetamide as a major component of human plasmatic metabolic profiling of benznidazole

Marson

Altcheh

Moscatelli

et al. 2014

Eur J Drug Metab Pharmacokinet

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Chagas disease is an endemic infection in Latin America with a high health impact. Caused by the parasite Trypanosoma cruzi, it has expanded to non-endemic regions such as North America and European countries via immigration of infected people. This infectious disease has been rising in the ranking of international health priorities due to the growing migration flows from endemic to non-endemic areas. Benznidazole (BZN), a nitroheterocyclic drug, is one of the two trypanocidal drugs currently in clinical use, associated with significant adverse drug reactions (ADRs). Mammalian metabolism of BNZ has been poorly studied, including the potential role of metabolites on both toxicity and anti-parasitic activity. High-resolution UPLC/MS/MS was used to analyze three plasma samples obtained from pediatric patients under BNZ treatment in steady state. Spectroscopic and structural criteria were applied to identify BNZ and accompanying substances from chromatographic signals. From all detected species, two can be undoubtedly associated with the BNZ and N-benzylacetamide molecules, the second one being a fragment of the parent drug (BZN). From the obtained results, two hypotheses could be formulated. The first one is to relate the presence of N-benzyl acetamide with the hepatic metabolism of BNZ. The second hypothesis has to do with the possible trypanocidal activity of this metabolite, as well as its role in the development of side effects, associated with the pharmacotherapy. Complementary studies should be carried out to determine the possible association of this metabolite with the BNZ treatment stages, patient's clinical features, ADRs, and trypanocidal effectiveness.

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“…The detection of this substance, although not identified, was previously reported by our work group (Marson et al 2013).…”

Section: Introductionsupporting

confidence: 56%

Section: Patient Plasma Samples and Pretreatmentmentioning

confidence: 99%

Identification of N-benzylacetamide as a major component of human plasmatic metabolic profiling of benznidazole

Marson

Altcheh

Moscatelli

et al. 2014

Eur J Drug Metab Pharmacokinet

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“…The LLOQ was established as 0.15 μg/mL, given the criteria of precision and accuracy (Table 2). This LLOQ was lower compared with other published methods, demonstrating that the proposed method was more sensitive for the quantification of plasma BNZ (Salomon, 2012;Maximiano et al, 2011;Morilla et al, 2002; Moreira da Silva et al, 2012; Leonardi et al, 2013;Marsón et al, 2013;Morilla et al, 2003). The recovery test also showed the acceptance criteria for precision and accuracy (Table 3), demonstrating that the method of processing the sample was suitable for the analysis of BNZ in plasma.…”

Section: Methods Validationmentioning

confidence: 57%

“…In some of these studies, bioanalytical methods were developed for the quantification of BNZ in plasma with application in vivo studies to evaluate the pharmacokinetics of the drug. These methods were developed using high-performance liquid chromatography (HPLC) systems with differences in solvent proportions of mobile phase, sample volume, run time, linearity, sensitivity, injection volume and sample preparation, among other factors (Morilla et al, 2002;Moreira da Silva et al, 2012;Leonardi et al, 2013;Marsón et al, 2013;Padró et al, 2013;Guerrero et al, 2011;Pinazo et al, 2013;Workman et al, 1984;White et al, 1982).…”

Section: Introductionmentioning

confidence: 99%

Rapid and sensitive ultra‐high‐pressure liquid chromatography method for quantification of antichagasic benznidazole in plasma: application in a preclinical pharmacokinetic study

Davanço

Campos

Peccinini

2014

Biomedical Chromatography

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Benznidazole (BNZ) and nifurtimox are the only drugs available for treating Chagas disease. In this work, we validated a bioanalytical method for the quantification of BNZ in plasma aimed at improving sensitivity and time of analysis compared with the assays already published. Furthermore, we demonstrated the application of the method in a preclinical pharmacokinetic study after administration of a single oral dose of BNZ in Wistar rats. A Waters® Acquity UHPLC system equipped with a UV-vis detector was employed. The method was established using an Acquity® UHPLC HSS SB C18 protected by an Acquity® UHPLC HSS SB C18 VanGuard guard column and detection at 324 nm. The mobile phase consisted of ultrapure water-acetonitrile (65:35), and elution was isocratic. The mobile phase flow rate was 0.55 mL/min, the volume of injection was 1 μL, and the run time was just 2 min. The samples were kept at 25°C until injection and the column at 45°C for the chromatographic separation. The sample preparation was performed by a rapid protein precipitation with acetonitrile. The linear concentration range was 0.15-20 µg/mL. The pharmacokinetic parameters of BNZ in rats were determined and the method was considered sensitive, fast and suitable for application in pharmacokinetic studies.

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“…However, several methods have been described for BNZ quantification in animal and human plasma, urine, and milk (12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22). To evaluate BNZ biodistribution, a bioanalytical method was developed to determine its quantification in mouse tissues, including spleen, brain, heart, colon, liver, lung, and kidney.…”

mentioning

confidence: 99%

Pharmacokinetics and Tissue Distribution of Benznidazole after Oral Administration in Mice

Perin

Silva

Fonseca

et al. 2017

Antimicrob Agents Chemother

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Specific chemotherapy using benznidazole (BNZ) for Chagas disease during the chronic stage is controversial due to its limited efficacy and toxic effects. Although BNZ has been used to treat Chagas disease since the 1970s, few studies about the biodistribution of this drug exist. In this study, BNZ tissue biodistribution in a murine model and its pharmacokinetic profile in plasma were monitored. A bioanalytical high-performance liquid chromatography method with a UV detector (HPLC-UV) was developed and validated according to the European Medicines Agency for quantification of BNZ in organs and plasma samples prepared by liquidliquid extraction using ethyl acetate. The developed method was linear in the BNZ concentration, which ranged from 0.1 to 100.0 g/ml for plasma, spleen, brain, colon, heart, lung, and kidney and from 0.2 to 100.0 g/ml for liver. Validation assays demonstrated good stability for BNZ under all conditions evaluated. Pharmacokinetic parameters confirmed rapid, but low, absorption of BNZ after oral administration. Biodistribution assays demonstrated different maximum concentrations in organs and similar times to maximum concentration and mean residence times, with means of 40 min and 2.5 h, respectively. Therefore, the biodistribution of BNZ is extensive, reaching organs such as the heart and colon, which are the most relevant organs affected by Trypanosoma cruzi infection, and also the spleen, brain, liver, lungs, and kidneys. Simultaneous analyses of tissues and plasma indicated high BNZ metabolism in the liver. Our results suggest that low bioavailability, instead of inadequate biodistribution, could be responsible for therapeutic failure during the chronic phase of Chagas disease.

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Development of UV/HPLC Methods for Quantitative Analysis of Benznidazole in Human Plasma and Urine for Application in Pediatric Clinical Studies

Abstract: The LOQ of methods seems appropriate in pediatric clinical contexts. Both procedures were applied with good results, to the quantization of BNZ in plasma and urine of patients treated for Chagas disease.

Cited by 16 publications

References 18 publications

Identification of N-benzylacetamide as a major component of human plasmatic metabolic profiling of benznidazole

Identification of N-benzylacetamide as a major component of human plasmatic metabolic profiling of benznidazole

Rapid and sensitive ultra‐high‐pressure liquid chromatography method for quantification of antichagasic benznidazole in plasma: application in a preclinical pharmacokinetic study

Pharmacokinetics and Tissue Distribution of Benznidazole after Oral Administration in Mice

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