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Analytical Procedures

Preclinical Monitoring of Drug Association in Experimental Chemotherapy of Chagas' Disease by a New HPLC-UV Method

Rodrigo Moreira da Silva, Líliam Teixeira Oliveira, Neila Márcia Silva Barcellos, Jacqueline de Souza, Marta de Lana
Rodrigo Moreira da Silva
aDepartamento de Farmácia, Programa de Pós-Graduação em Ciências Farmacêuticas (CiPHARMA), Escola de Farmácia, Universidade Federal de Ouro Preto, Ouro Preto, Brazil
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Líliam Teixeira Oliveira
aDepartamento de Farmácia, Programa de Pós-Graduação em Ciências Farmacêuticas (CiPHARMA), Escola de Farmácia, Universidade Federal de Ouro Preto, Ouro Preto, Brazil
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Neila Márcia Silva Barcellos
aDepartamento de Farmácia, Programa de Pós-Graduação em Ciências Farmacêuticas (CiPHARMA), Escola de Farmácia, Universidade Federal de Ouro Preto, Ouro Preto, Brazil
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Jacqueline de Souza
aDepartamento de Farmácia, Programa de Pós-Graduação em Ciências Farmacêuticas (CiPHARMA), Escola de Farmácia, Universidade Federal de Ouro Preto, Ouro Preto, Brazil
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Marta de Lana
bDepartamento de Análises Clínicas, Programa de Pós-Graduação em Ciências Farmacêuticas (CiPHARMA), Escola de Farmácia and Programa de Pós-Graduação em Ciências Biológicas (NUPEB), Universidade Federal de Ouro Preto, Ouro Preto, Brazil
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DOI: 10.1128/AAC.05785-11
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    Fig 1

    (A) Mean plasma concentration profiles of BNZ administered alone and in association with ITC in mice (n = 5) after a single oral dose of 5 mg of each drug. (B) Highlight of the final portion of the curve displayed in panel A.

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    Fig 2

    Mean plasma concentration profiles of ITC administered alone and in association with BNZ in mice (n = 5) after a single oral dose of 5 mg of each drug.

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  • Table 1

    Validation parameters of the HPLC-UV method developed for simultaneous quantification of BNZ and ITC in plasma of mice (mean, 5 repetitions)

    Validation parameterValue
    BNZITC
    Sensitivity
        Limit of quantification (μg/ml)0.51.0
    Linearity
        Correlation coefficient0.99850.9924
        Mean regression equationy = 0.0522x + 0.0229y = 0.0128x + 0.0111
    Reproducibility
        Mean accuracy (%)b96.6793.37
        Mean precision (RSD) (%)a3.875.43
    Recovery
        Mean recovery (%)92.4693.92
    • ↵a Mean precision of intra- and interday analyses; RSD, relative standard deviation.

    • ↵b Mean accuracy of intra- and interday analyses.

  • Table 2

    Pharmacokinetic parameters of BNZ and ITC administered alone and in association in mice after a single oral dose of 5 mg of each drug

    ParameterValuea when administered:
    AloneIn association
    BNZITCBNZITC
    Cmax (μg/kg)151.28 ± 6.03b11.76 ± 1.33101.19 ± 5.10b10.55 ± 0.96
    Tmax (h)2.2 ± 0.24.6 ± 0.242.4 ± 0.245.0 ± 0.32
    AUC0-t (μg · h/ml)539.14 ± 14.5465.93 ± 2.99612.33 ± 50.8777.45 ± 9.17
    AUC0-∞ (μg · h/ml)547.53 ± 11.9679.51 ± 4.69673.90 ± 64.2494.21 ± 12.61
    AUMC0-t (μg · h2/ml)2,635.39 ± 65.66b526.67 ± 41.657,465.97 ± 1,345.0b949.05 ± 231.0
    AUMC0-∞ (μg · h2/ml)2,757.49 ± 86.19b753.16 ± 74.5310,817.19 ± 2,112.0b1,377.09 ± 386.6
    MRT (h)5.05 ± 0.22b9.38 ± 0.4215.47 ± 1.7b13.52 ± 2.17
    CL (ml/h/kg)416.74 ± 10.923,417.30 ± 149.50375.2 ± 28.93,077.97 ± 404.4
    V (ml/kg)2,112.33 ± 136.90b31,870.60 ± 970.805,621.29 ± 209.5b38,710.66 ± 2,604.0
    λz (1/h)0.51 ± 0.09b0.23 ± 0.040.05 ± 0.00b0.21 ± 0.04
    t½β (h)1.64 ± 0.41b3.39 ± 0.4712.18 ± 0.78b3.89 ± 0.79
    • ↵a Mean ± SE of five experiments.

    • ↵b Significant differences in each experimental group at P < 0.05.

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Preclinical Monitoring of Drug Association in Experimental Chemotherapy of Chagas' Disease by a New HPLC-UV Method
Rodrigo Moreira da Silva, Líliam Teixeira Oliveira, Neila Márcia Silva Barcellos, Jacqueline de Souza, Marta de Lana
Antimicrobial Agents and Chemotherapy May 2012, 56 (6) 3344-3348; DOI: 10.1128/AAC.05785-11

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Preclinical Monitoring of Drug Association in Experimental Chemotherapy of Chagas' Disease by a New HPLC-UV Method
Rodrigo Moreira da Silva, Líliam Teixeira Oliveira, Neila Márcia Silva Barcellos, Jacqueline de Souza, Marta de Lana
Antimicrobial Agents and Chemotherapy May 2012, 56 (6) 3344-3348; DOI: 10.1128/AAC.05785-11
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