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Simultaneous quantitation of chloroquine and primaquine by UPLC-DAD and comparison with a HPLC-DAD method.

Miranda TA, Silva PH, Pianetti GA, César IC - Malar. J. (2015)

Bottom Line: UPLC method was fully validated and the results were compared to a conventional HPLC-DAD method for the analysis of chloroquine and primaquine in tablet formulations.No significant differences were observed between the chloroquine and primaquine contents obtained by UPLC and HPLC methods.The developed UPLC method was shown to be a rapid and suitable technique to quantify chloroquine and primaquine in pharmaceutical preparations and may be successfully employed for quality control analysis.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Produtos Farmacêuticos, Faculdade de Farmácia, Universidade Federal de Minas Gerais, Av Pres Antônio Carlos 6627, Belo Horizonte, 31270-901, MG, Brazil. isaccesar@bol.com.br.

ABSTRACT

Background: Chloroquine and primaquine are the first-line treatment recommended by World Health Organization for malaria caused by Plasmodium vivax. Since the problem of counterfeit or substandard anti-malarials is well established all over the world, the development of rapid and reliable methods for quality control analysis of these drugs is essential. Thus, the aim of this study was to develop and validate a novel UPLC-DAD method for simultaneously quantifying chloroquine and primaquine in tablet formulations.

Methods: The UPLC separation was carried out using a Hypersil C18 column (50 × 2.1 mm id; 1.9 μm particle size) and a mobile phase composed of acetonitrile (A) and 0.1% aqueous triethylamine, pH 3.0 adjusted with phosphoric acid (B), at a flow rate 0.6 mL/min. Gradient elution was employed. UV detection was performed at 260 nm. UPLC method was fully validated and the results were compared to a conventional HPLC-DAD method for the analysis of chloroquine and primaquine in tablet formulations.

Results: UPLC method was shown to be linear (r2 > 0.99), precise (CV < 2.0%), accurate (recovery rates from 98.11 to 99.83%), specific, and robust. No significant differences were observed between the chloroquine and primaquine contents obtained by UPLC and HPLC methods. However, UPLC method promoted faster analyses, better chromatographic performance and lower solvent consumption.

Conclusions: The developed UPLC method was shown to be a rapid and suitable technique to quantify chloroquine and primaquine in pharmaceutical preparations and may be successfully employed for quality control analysis.

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Related in: MedlinePlus

Chromatograms obtained by UPLC (full line) and HPLC (dotted line) for analysis of chloroquine (CHLO) and primaquine (PRIM) in tablet formulations.
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Fig2: Chromatograms obtained by UPLC (full line) and HPLC (dotted line) for analysis of chloroquine (CHLO) and primaquine (PRIM) in tablet formulations.

Mentions: Chromatographic parameters employed for the UPLC method development were initially adapted from HPLC method already used for routine analysis in our laboratory. The chromatographic parameters were firstly evaluated using C18 column and isocratic elution with a mobile phase composed of 60% organic solvent. Under these conditions, the retention factor obtained for chloroquine was considerably low. However, the decrease of organic solvent percentage in mobile phase increased the retention of primaquine peak, leading to a long run time, besides causing peak tailing. The use of gradient elution promoted adequate retention times and peak shapes for both anti-malarial drugs. Acetonitrile instead of methanol showed to be a better choice to assure the adequate separation of peaks. Finally, different pH values of mobile phase were evaluated and better peak symmetries were obtained at pH 3.0. Under these optimized conditions, chloroquine and primaquine peaks eluted at 1.0 and 1.3 min, respectively (Figure 2).Figure 2


Simultaneous quantitation of chloroquine and primaquine by UPLC-DAD and comparison with a HPLC-DAD method.

Miranda TA, Silva PH, Pianetti GA, César IC - Malar. J. (2015)

Chromatograms obtained by UPLC (full line) and HPLC (dotted line) for analysis of chloroquine (CHLO) and primaquine (PRIM) in tablet formulations.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4318193&req=5

Fig2: Chromatograms obtained by UPLC (full line) and HPLC (dotted line) for analysis of chloroquine (CHLO) and primaquine (PRIM) in tablet formulations.
Mentions: Chromatographic parameters employed for the UPLC method development were initially adapted from HPLC method already used for routine analysis in our laboratory. The chromatographic parameters were firstly evaluated using C18 column and isocratic elution with a mobile phase composed of 60% organic solvent. Under these conditions, the retention factor obtained for chloroquine was considerably low. However, the decrease of organic solvent percentage in mobile phase increased the retention of primaquine peak, leading to a long run time, besides causing peak tailing. The use of gradient elution promoted adequate retention times and peak shapes for both anti-malarial drugs. Acetonitrile instead of methanol showed to be a better choice to assure the adequate separation of peaks. Finally, different pH values of mobile phase were evaluated and better peak symmetries were obtained at pH 3.0. Under these optimized conditions, chloroquine and primaquine peaks eluted at 1.0 and 1.3 min, respectively (Figure 2).Figure 2

Bottom Line: UPLC method was fully validated and the results were compared to a conventional HPLC-DAD method for the analysis of chloroquine and primaquine in tablet formulations.No significant differences were observed between the chloroquine and primaquine contents obtained by UPLC and HPLC methods.The developed UPLC method was shown to be a rapid and suitable technique to quantify chloroquine and primaquine in pharmaceutical preparations and may be successfully employed for quality control analysis.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Produtos Farmacêuticos, Faculdade de Farmácia, Universidade Federal de Minas Gerais, Av Pres Antônio Carlos 6627, Belo Horizonte, 31270-901, MG, Brazil. isaccesar@bol.com.br.

ABSTRACT

Background: Chloroquine and primaquine are the first-line treatment recommended by World Health Organization for malaria caused by Plasmodium vivax. Since the problem of counterfeit or substandard anti-malarials is well established all over the world, the development of rapid and reliable methods for quality control analysis of these drugs is essential. Thus, the aim of this study was to develop and validate a novel UPLC-DAD method for simultaneously quantifying chloroquine and primaquine in tablet formulations.

Methods: The UPLC separation was carried out using a Hypersil C18 column (50 × 2.1 mm id; 1.9 μm particle size) and a mobile phase composed of acetonitrile (A) and 0.1% aqueous triethylamine, pH 3.0 adjusted with phosphoric acid (B), at a flow rate 0.6 mL/min. Gradient elution was employed. UV detection was performed at 260 nm. UPLC method was fully validated and the results were compared to a conventional HPLC-DAD method for the analysis of chloroquine and primaquine in tablet formulations.

Results: UPLC method was shown to be linear (r2 > 0.99), precise (CV < 2.0%), accurate (recovery rates from 98.11 to 99.83%), specific, and robust. No significant differences were observed between the chloroquine and primaquine contents obtained by UPLC and HPLC methods. However, UPLC method promoted faster analyses, better chromatographic performance and lower solvent consumption.

Conclusions: The developed UPLC method was shown to be a rapid and suitable technique to quantify chloroquine and primaquine in pharmaceutical preparations and may be successfully employed for quality control analysis.

Show MeSH
Related in: MedlinePlus