Chromatography fractions from the ethyl acetate fraction of Harungana madagascariensis Lam. Ex Poiret fruits (Hypericaceae) inhibited the growth of Mycobacterium tuberculosis

O. Afieroho, Onyinye Blessing Okonkwo, J. Bimba, T. T. Eliya, Augustina Uche Osuji, K. Abo
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Abstract

The economic burden of drug resistant tuberculosis strain is enormous due to its being one of the leading cause of global mortality. New treatments that are faster, simpler and affordable are urgently needed. Thus this research aimed to validate that the ethyl acetate fraction from the Nigeria variety of Harungana madagascariensis contain secondary metabolites with anti-Mycobacterium tuberculosis activity. The fruit of H. madagascariensis was extracted with 70% aqueous ethanol by cold maceration. The crude aqueous ethanol extract was defatted with n-hexane by partitioning and the aqueous portion further partitioned with ethyl acetate to give ethyl acetate portion (EAP) used in this study. Fractionation of EAP was done using chromatographic techniques. Phytochemical screening of the fractions was done using standard methods. Anti- Mycobacterium tuberculosis screening was done using Lowestein Jensen(LJ) method (test fractions stock solution 1mg/ml diluted to a 0.01mg/ml in the LJ medium while isoniazid, dihydrostreptomycin, ethambutol and rifampicin at their reported minimum inhibition concentrations(MIC) were used as reference anti-TB drugs). Four (4) chromatography fractions (EAF1 –EAF4) were obtained. All fractions were found to contain phenolics with anthraquinone present in EAF2 and EAF3. Fractions EAF3 and EAF4 inhibited the growth of the Mycobacteria tuberculosis. This preliminary screening validates the use of H. madagascariensis in traditional medicine and as a potential source of natural products with anti-Mycobacterium tuberculosis activity as well as a good source of anthraquinone.
马达加斯加小檗乙酸乙酯部分的层析组分。金丝桃科水果对结核分枝杆菌的生长有抑制作用
耐药结核菌株的经济负担是巨大的,因为它是全球死亡的主要原因之一。我们迫切需要更快、更简单、负担得起的新疗法。因此,本研究旨在验证尼日利亚品种马达加斯加哈隆加纳的乙酸乙酯部分含有具有抗结核分枝杆菌活性的次级代谢物。用70%乙醇冷浸法提取马达加斯加红桃果实。将粗乙醇水提物用正己烷分馏脱脂,再用乙酸乙酯分馏得到本研究用的乙酸乙酯部分(EAP)。用色谱技术对EAP进行分离。采用标准方法进行植物化学筛选。采用Lowestein Jensen(LJ)法进行抗结核分枝杆菌筛查(LJ培养基中1mg/ml原液稀释至0.01mg/ml,异烟肼、二氢链霉素、乙胺丁醇和利福平以其报道的最低抑制浓度(MIC)作为抗结核参比药物)。得到EAF1 -EAF4 4个层析组分。EAF2和EAF3均含有蒽醌类酚类物质。EAF3和EAF4组分抑制结核分枝杆菌的生长。这一初步筛选验证了马达加斯加分枝杆菌在传统医学中的应用,以及它作为具有抗结核分枝杆菌活性的天然产物的潜在来源以及蒽醌的良好来源。
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