Development of ELISA-based methods to measure the anti-malarial drug chloroquine in plasma and in pharmaceutical formulations

Malaria Journal

Methodology

Camilla Recke¹  Ib C Bygbjerg²  Insaf F Khalil²  Michael Alifrangis²  Lotte C Hoegberg³  Claus Koch⁴  Anita Ronn⁵ 

[1] Bio Port A/S/, Grusbakken 8, DK-2820, Gentofte, Denmark;Centre for Medical Parasitology at Department of International Health, Immunology and Microbiology, University of Copenhagen and Department of Infectious Diseases, Copenhagen University Hospital, CSS Oster Farimagsgade 5, 1014, Copenhagen K, Denmark;Department of Toxicology, Bispebjerg Hospital, Bispebjerg Bakke, 23 2400, Copenhagen NV, Denmark;Institute of molecular medicine, Department of Cancer and Inflammation, University of Southern Denmark, DK-5000, Odense, Denmark;null;

关键词: High Performance Liquid Chromatography;    Malaria;    Chloroquine;    Pharmaceutical Formulation;    Vivax Malaria;   

DOI  :  10.1186/1475-2875-10-249

 received in 2011-02-06, accepted in 2011-08-24,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundIn Central and South America and Eastern and Southern Africa, Plasmodium vivax infections accounts for 71-81% and 5% of malaria cases, respectively. In these areas, chloroquine (CQ) remains the treatment of choice for P. vivax malaria. In addition, CQ has recently proven to be an effective HIV-1 therapeutic agent. There is a dire need to continue monitoring quality of CQ as there is a major influx of substandard and fake formulations into malaria-endemic countries. The use of fake/substandard drugs will result in sub-therapeutic levels endangering the patient and possibly select for parasite resistance. The aim of this study was to develop an inexpensive, simple antibody-based ELISA to measure CQ concentrations in tablets and in plasma.MethodsA monoclonal antibody (MAb) that reacts with the N-side chain of the CQ molecule was prepared by use of a CQ analogue. A specific and reliable ELISA for detection of CQ was developed. The developed assay was validated by measuring CQ in tablets sold in Denmark, India and Sudan. Furthermore, kinetics of CQ concentrations in plasma of four volunteers, who ingested two tablets of Malarex® containing, 250 mg CQ base, were measured before drug intake, three hours later and thereafter at days 1, 3, 7, 14, 21 and 28. The same plasma samples were simultaneously measured by high performance liquid chromatography (HPLC).ResultsThe ELISA proved an easy-to-handle and very sensitive tool for the detection of CQ with a lower limit of detection at 3.9 ng/ml. ELISA levels of CQ in plasma showed high agreement with the levels obtained by HPLC (r = 0.98). The specificity in the negative control group was 100%.ConclusionThe developed ELISA can be used for quality screening of CQ in pharmaceutical formulations and for drug monitoring in malaria and in other infectious diseases, such as HIV, where CQ proved to be an effective therapeutic agent. The methodology has been exploited to develop monoclonal antibodies for the drugs used in artemisinin-based combination therapy (ACT).

【 授权许可】

CC BY   
© Khalil et al; licensee BioMed Central Ltd. 2011