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01.12.2012 | Methodology | Ausgabe 1/2012 Open Access

Malaria Journal 1/2012

A simple and inexpensive haemozoin-based colorimetric method to evaluate anti-malarial drug activity

Malaria Journal > Ausgabe 1/2012
Tran Thanh Men, Nguyen Tien Huy, Dai Thi Xuan Trang, Mohammed Nasir Shuaibu, Kenji Hirayama, Kaeko Kamei
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​1475-2875-11-272) contains supplementary material, which is available to authorized users.

Competing interests

The authors declare that they have no competing interests.

Authors’ contributions

NTH and KK developed the idea for the project. TTM and NTH conceived and designed the experiments. TTM, NTH and MNS carried out the laboratory work. TTM, NTH, DTXT, KH, and KK analysed and interpreted the data. TTM, NTH, MNS, KH and KK contributed reagents/materials/analysis tools. . TTM, NTH, DTXT and KK wrote the paper. All authors had full access to all data in the study, read and approved the manuscript.



The spread of drug resistance in malaria parasites and the limited number of effective drugs for treatment indicates the need for new anti-malarial compounds. Current assays evaluating drugs against Plasmodium falciparum require expensive materials and equipment, thus limiting the search for new drugs, particularly in developing countries. This study describes an inexpensive procedure that is based on the advantage of a positive correlation between the haemozoin level of infected erythrocytes and parasite load.


The relationship between parasitaemia and the haemozoin level of infected erythrocytes was investigated after converting haemozoin into monomeric haem. The 50% inhibitory concentration (IC50) values of chloroquine, quinine, artemisinin, quinidine and clotrimazole against P. falciparum K1 and 9A strains were determined using the novel assay method.


The haemozoin of parasites was extracted and converted into monomeric haem, allowing the use of a colorimeter to efficiently and rapidly measure the growth of the parasites. There was a strong and direct linear relationship between the absorbance of haem converted from haemozoin and the percentage of the parasite (R2 = 0.9929). Furthermore, the IC50 values of drugs were within the range of the values previously reported.


The haemozoin-based colorimetric assay can be considered as an alternative, simple, robust, inexpensive and convenient method, making it applicable in developing countries.
Authors’ original file for figure 1
Authors’ original file for figure 2
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