Comparative Clinical Study of Mist Amen Fevermix and Edhec Malacure: Two Polyherbal Products used for the Treatment of uncomplicated malaria in Ghana against Artemether/Lumefantrine

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2020-11
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The use of herbal medicinal products for the treatment of malaria an infectious and a life threatening disease, has increased globally. However, inadequate scientific studies, questions about the quality, safety and efficacy of such herbal products have been raised. On the other hand, the reduced sensitivity of the malaria parasites to artemisinin-based combination therapies is also of concern. There is therefore the need for new antimalarial medications including those from alternative sources such as herbal medicinal products. In this study, methods for the quality control of Mist Amen Fevermix and Edhec Malacure, two polyherbal antimalarial products used in Ghana for the management of uncomplicated malaria was undertaken. The development of the quality parameters for the test samples was based on phytochemical, physicochemical, chromatographic and spectroscopic methods. The set parameters were found to be sufficient to evaluate Mist Amen Fevermix and Edhec Malacure, and can be used as reference standards for the quality control purposes. Qualitative phytochemical screening and fingerprinting were undertaken based on standard analytical methods. The antiplasmodial activity was assessed in vitro by using field isolates of Plasmodium falciparum with SYBR® Green assays to measure parasite growth inhibition. Thermo Elemental M5 Atomic Absorption Spectrophotometer (AAS) fitted with Graphite furnace and an auto sampler was used to determine the heavy metal contents of the herbal products. The herbal samples were evaluated for microbial load by using the appropriate culture media. In vivo antiparasitic activity in mice was assessed using the Rane’s curative method using ANKA strain of Plasmodium berghei parasites. A comparative clinical study was done to assess the safety and effectiveness of the test samples at the Tafo Government Hospital, Kumasi after Committee on Human Research, Publication and Ethics approval. Male and female patients aged 15-45 years with clinically established malaria were treated with Mist Amen Fevermix and Edhec Malacure, at the specified doses of 45 mls (0.1063 g) and 30 mls (0.0521 g) three times daily after meals for three days. Basic phytochemical screening of the two products indicated the presence of the following phytochemicals: alkaloids, saponins, tannins, phytosterols and flavonoids. From the data, it was established that Mist Amen Fevermix and Edhec Malacure complied with the pharmacopoeial standards after testing for microbes. The following heavy metals were present in Mist Amen Fevermix and Edhec Malacure: Fe, Ni, K, Zn, Hg, Cu, Mn, Cr, Cd, Pb, Fe, Cu, K and Na. Ni was below detectable limit in Edhec Malacure. The phytochemical screening of the products revealed the presence of alkaloid flavonoid, tannin, steroid and saponin. The HPLC method was validated for linearity, limits of detection and quantification, precision and accuracy. The test products were found not to have been adulterated with lumefantrine, artemether and quinine. The test herbal products showed in vitro and in vivo antiplasmodial activities against Plasmodium falciparum and Plasmodium berghei parasites. Inhibitory concentration (IC50) values for Edhec Malacure was 70.89 ng/ml and that of Mist Amen Fevermix was 112.5 ng/ml. Edhec Malacure suppressed 76.17% of parasitaemia while Mist Amen Fevermix suppressed 69.03% of parasitaemia. Edhec Malacure demonstrated curative chemo suppressive potentials of 80.93% at the dose of 2.234 mgkg-1 and Mist Amen Fevermix % suppression was 69.03% at a dose of 4.56mg/kg-1. Both products demonstrated antiplasmodial activity in human red blood cells. The clinical evaluation of the test samples showed that Mist Amen Fevermix exhibited a statistically significant difference between the mean malaria parasite load recorded at the first visit and those recorded at the second visit, t(23) = 4.59, p =0 .000. Similarly, there was a significant difference between the mean parasite count recorded on the second visit and the third visit, t(6) = 1.49, p =0 .187. No difference were recorded for the third and fourth visits t(3) = 1.00, p =0 .391. Edhec Malacure also exhibited a significant difference in efficacy between the mean malaria parasite count recorded at the first visit and those recorded at the second visit, t(26) =3.77, p =0 .001. Similarly, there is a statistically significant difference between malaria parasite count at the second visits and third visits, t(16) = 1.74, p =0 .100. This shows the significant effectiveness of the products. Kidney and liver panel as well as full blood count and vital signs were within normalviii reference range at the end of the 28-day study and thus established the safety of Mist Amen Fevermix and Edhec Malacure in the treatment of uncomplicated malaria. The results support claims that Mist Amen Fevermix and Edhec Malacure may be useful antimalarial agents. This study has demonstrated the in vitro and in vivo antiplasmodial activities of Mist Amen Fevermix and Edhec Malacure, and suggests that, the products have promising antimalarial activity. The in vivo findings showed that Mist Amen Fevermix and Edhec Malacure are relatively safe for oral administration at doses tested. In addition, the study supports the use of Mist Amen Fevermix and Edhec Malacure, two polyherbal products for the treatment of uncomplicated malaria. Both products achieved a comparable clinical treatment outcome to the reference control medication artemether/lumefantrine.
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A thesis submitted to the Department of Pharmacognosy, Kwame Nkrumah University of Science and Technology, Kumasi in partial fulfilment of the requirements for the award degree of DOCTOR OF PHILOSOPHY IN PHARMACOGNOSY
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