Desenvolvimento e validação de métodos analíticos para quantificação de cloroquina e primaquina em comprimidos e em plasma humano

• Ano de defesa: 2013
• Autor(a) principal: Tiago Assis Miranda
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Minas Gerais; UFMG
• Palavras-chave em Português: espectrometria de massas; cloroquina; CLAE; plasma humano; CLUE; antimaláricos; primaquina; Validação de método; Cromatografia líquida de alta eficiência; Tecnologia farmaceutica; Cloroquina; Espectrometria de massa; Antimaláricos
• Link de acesso: http://hdl.handle.net/1843/EMCO-9MVFUB

Malaria is a disease that has affected humanity for a long time, it is believed that 3.3 billion people live in areas at risk of malaria infection. Although declining, the epidemiological situation of malaria in Brazil is worrying today. In 2011, 267 045 cases were registered in the country, of which 87% caused by Plasmodium vivax. In the treatment of malaria by P. vivax, are used chloroquine and primaquine, according to the Ministry of Health. It is extremely important to know the pharmacokinetics of chloroquine and primaquine, when administered simultaneously, in order to guarantee that during treatment, both are above the minimum plasma concentration necessary for pharmacological action. Concomitant to the needs of antimalarial therapeutic monitoring, development and application of analytical methods for identification and quantification of these drugs are of great importance to evaluate and ensure the quality of drugs currently marketed and distributed. The aim of this study was to develop and validate analytical methods for the simultaneous quantification of chloroquine and primaquine tablets and in human plasma. To determine the drugs into tablets, two analytical methods have been developed and validated: high performance liquid chromatography (HPLC) and ultrahigh performance liquid chromatography (UHPLC). The HPLC analysis was performed in a C18 column (100 x 4.6 mm; 5 ìm) and mobile phase composed of acetonitrile and 0.1% (v/v) triethylamines solution with pH adjusted to 3.0 with phosphoric acid, in gradient mode. The flow rate was 1000 ìL/min and the injection volume was 10 ìL. The UV detection was in 260 nm. The UHPLC analysis was performed in a C18 column (50 x 2.1 mm; 1.9 ìm) under the same conditions HPLC method, except for the flow rate was 600 ìL/min and the injection volume was 7 ìL. The developed method complied with all required validation parameters and showed to be adequate for routine analysis. A bioanalytical method by HPLC with detection by mass spectrometry and eletrospray ionization in the positive mode was developed for the quantitation of chloroquine and primaquine in human plasma, with amodiaquine selected as an internal standard. For the sample preparation, was selected to liquid-liquid extraction. The chromatographic analysis was performed using a cyano column (150 x 4.6 mm; 5 ìm) and the mobile phase composed of methanol and 0.01 M ammonium acetate solution with pH adjusted to 3.0 with formic acid, in isocratic mode. The flow rate was 1000 ìL/min and the injection volume was 20 ìL. The employed transitions for the quantitation of chloroquine, primaquine and amodiaquine were m/z 320,49 m/z 246,94, m/z 260,48 m/z 174,88 and m/z 356,59 m/z 282,97, respectively. The method proved to be selective and provides recovery rates close to 100% for all drugs.