Abstract
Aim: The present study aims to investigate the phytochemical, antimicrobial and acute toxicity assay of methanol extract of Citrus medica L. fruit (CMF) and Citrus hystrix D.C. fruit (CHF).
 Place and Duration of Study: Fruit samples were collected between February to August 2018, at the Department of Life Sciences, Manipur University.
 Methodology: Phytochemical studies were conducted using Gas Chromatography-Mass Spectrometry (GC-MS), HR-LC-MS (High Resolution-Liquid Chromatography-Mass Spectrometry), Graphite Furnace-Atomic Absorption Spectrometry (GF-AAS), and Inductively Coupled Plasma-Optical Emission Spectroscopy (ICP-OES) respectively. The standard filtered disc-diffusion method was used for antimicrobial assay. Acute toxicity was performed using 423-OECD guidelines.
 Results: GC-MS and HR-LC-MS analysis showed presence of Ranitidine, 4-Methylesculetin, Diosmin and Avobenzone in CMF whereas 9-Octadecenamide, Gamma-Sitosterol, n-Hexadecanoic acid, 2-Methoxy-4-Vinylphenol, Rhoifolin, Diosmin and Phytosphingosine in CHF. GF-AAS and ICP-OES study prominently showed Pb content in both the samples. Highest element in CMF was Pb (4.26±0.120 ppm) while in CHF was Cr (4.35±0.70 ppm). Antimicrobial study exhibited highest inhibitory effect of CMF against Staphylococcus aureus and Klebsiella pneumonia while CHF against Escherichia coli, Klebsiella pneumonia and Staphylococcus aureus than Gentamicin (p<0.05). No toxicity behaviour and mortality in mice were observed during acute toxicity study period even at a dose of 5000 mg/kg body weight. Changes in serum constituent level were observed however, no genotoxicity was recorded.
 Conclusion: We concluded that CMF and CHF cultivation site selection should be the first step to avoid Pb content. The CMF and CHF have many health beneficial constituents. From this study also concluded that CMF and CHF may be a potential source of antiulcer, antimicrobial, antiarthritic, diuretic, antiinflammatory and anticancer effects. However, further study to understand whether changes in serum constituent level for prolonged period usages as medicine or nutraceuticals is highly recommended.
Highlights
Manipur is heavily endowed with abundant natural resources and is home to millions of medicinal plants
The HR-LC-MS result of the Citrus medica L. fruit (CMF) extract showed 12 metabolites while the Citrus hystrix D.C. fruit (CHF) extract showed 9 metabolites with DB diff ranging between -0.1 to +1.58
Ranitidine and Diosmin had an inhibitory effect on gastric acid secretion [22] and protected against gastric injury [23]
Summary
Manipur is heavily endowed with abundant natural resources and is home to millions of medicinal plants. Various plants, including those used by the traditional medical practitioner, grow luxuriantly in Manipur, North-East India, within the Indo-Burmese mega-biodiversity hot-spot. The numerous medicinal plants used in Manipur include a variety of Citrus species. The taste of the Citrus fruits found in Manipur ranges from sweet to sour (acidic); some are generally consumed as edible fruit, while some are well known for their use in traditional medicine [5]. Among the varieties of Citrus, the fruit of Citrus medica L., known as Citron, is widely used to treat urolithiasis, especially calcium oxalate stone [8]
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