This article was posted by Dr. P.T. Ezhil Selvan BIOACTIVE COMPOUNDS FROM RHIZOMES OF ALPINIA OFFICINARUM HANCE INTRODUCTION The Siddha system of medicine is one of the ancient systems contemporaneous with those of the submerged lands, Egyptian, Mesopotamian, Chinese and Grecian medicines. The unique nature of this system is its continuous service to humanity for more than five thousand years in combating diseases and in maintaining physical, mental and moral health.1 Siddha medical system has its literature in Tamil and it is practiced in Tamil speaking parts of India. Its principles and doctrines, both fundamental and applied, have a close similarity to Ayurveda, with specialization in Iatrochemistry.2 According to a report by WHO - The Siddha system is capable of treating all types of disease other than emergency cases. In general this system is effective in treating all types of skin problems particularly Psoriasis, STD, urinary tract infections, diseases of liver and gastro intestinal tract, general debility, postpartum anaemia, diarrhoea and general fevers in addition to arthritis and allergic disorders.2 A report by WHO states, “Indian government’s research in the homoeopathic and the Siddha systems have been encouraging. Some of the medicines in these systems have the potential of reducing the viral (HIV) load in the body of the patients thus ensuring a healthier and longer life with the infection”.3 Plants are very efficient in synthesizing organic compounds via photosynthesis from inorganic materials found in the environment. The pathways for generally modifying and synthesizing carbohydrates, proteins, fats and nucleic acids are found to be essentially the same in all organisms. These processes are collectively described as primary metabolism, with the compound involved in the pathways being termed primary metabolites.4 In contrast to these primary metabolic pathways, there also exists an area of metabolism concerned with compounds which have a much more limited distribution in nature. Such compounds, called secondary metabolites, are found in only specific organisms or groups of organisms and are an expression of the individuality of species. Secondary metabolites are not necessarily produced under all conditions.4 It is this area of secondary metabolism that provides most of the pharmacologically active natural products. Some of the important secondary metabolites are alkaloids, flavonoids, terpenoids, glycosides, phenols, phenazines, polyketides, fatty acid, nonribosomal peptides and ribosomal peptides.5 In a review with 76 references, Zingiberaceae family contains 45 genera and over 750 species that are distributed in the tropical region, chiefly in Indo-Malaysia. In India, this family is represented by about 17 genera and 112 species occurring chiefly in Eastern Himalayas and Western Ghats. The best known examples of the family are ginger (Zingiber officinale), turmeric (Curcuma longa) and cardamom (Elettaria cardamomum).6 The plants of the family Zingiberaceae are widely used world over as food flavors, spices and for the medicinal purposes from the ancient time. Alpinia species are used in the oriental part of medicine and food additives, spices and in indigenous system of medicine. The various parts of Alpinia species are used in Indian systems of medicine and as food flavor. It is also used for the treatment of some inflammatory conditions, as an anticancer and anticalculi agent, as a digestive, spleen and liver tonic, and in dyspepsia, gastralgia, sea sickness and abdominal colic. Recent studies confirmed that Alpinia species possesses significant antiulcer, anticalculi, and anti-inflammatory activities.6 Alpinia officinarum Hance, which is called as Lesser Galangal of the family Zingiberaceae has been traditionally used in the Siddha system of medicine. The rhizome of the herb is used medicinally. The rhizome has an aromatic odour and spicy to taste. The rhizome is used for rheumatic disorders, cold, cough, fever, etc. It has been proved that it has very good activity against many bacteria, fungi, virus and many other pathogens.7 MATERIALS AND METHODS Collection and authentication of plant material: The dried rhizomes of Alpinia officinarum were procured from authorized suppliers from Chennai. The identity and authenticity of the Alpinia officinarum rhizomes was confirmed at Centre for Advanced Studies in Botany, University of Madras. Voucher specimens of Alpinia officinarum was deposited in CAS in botany herbarium. Extraction of active compound using Ethanol: The plant material was shade dried at room temperature. 100 grams of the dried Alpinia officinarum rhizome was powdered in a pulveriser. The ethanolic extract was taken by cold storage process where the cycles were repeated by adding fresh ethanol to the extract. The extract was concentrated and slurry is then filtered by what man filter paper No. 1 and dried. The 100mg of extract was dissolved in 1 ml of ethanol and made upto 100 ppm. TLC analysis of crude extracts of Alpinia officinarum: Crude ethanolic extracts of Alpinia officinarum was analyzed using Thin Layer Chromatography methods. Ideal solvent concentration was optimized for better elution of active principle. The following solvent combinations were tested to optimize the better elution of the active compounds from the crude and the respective Rf values were calculated. The ethanolic extract was separated by Thin Layer Chromatography on Silica Gel G (Merck precoated TLC plates) using different solvent systems; Benzene: Methanol (1: 2), Methanol: Benzene (1: 2), Acetone: Benzene (1: 1), Acetone: Methanol, (1: 1), Toluene: Methanol (1: 2), Methanol: Toluene (1: 2), Ethyl acetate: Methanol (2: 1), Benzene: Methanol: Chloroform (1: 1: 1), Acetone: Methanol: Chloroform (1: 1: 1), Toluene: Methanol: Chloroform (1: 1: 1), Benzene: Methanol: Toluene (1: 2: 1), Benzene: Methanol: Toluene: Chloroform (0.5: 2: 0.5: 1). After each separation, the TLC plates were exposed to iodine vapour in an iodine chamber to visualize the spots. Determination of antibacterial activity of Ethanolic extract: Disc preparation: The ethanol extract was used in disc diffusion assay to test for antibacterial activity. The disc was prepared using sterile disk (Himedia) 5 mm in diameter. The ethanol extracts were loaded to the each sterile disk, under a laminar flow at the increasing concentration in micro litter / milligram of these extracts was 30µl/3mg, 60µl/6mg and 90µl/9mg respectively were allowed to dry. Selection of bacterial strains for antimicrobial activity testing: Four bacterial test strains were selected namely, proteus sp., Staphylococcus aureus, Escherichia coli and Salmonella sp. The cultures were obtained from the culture collection of Centre for Advanced Studies in Botany, University of Madras. The 18 hours incubated inoculums were used for antimicrobial activity testing. Antimicrobial susceptibility test by disc diffusion method: The Kirby-Bauer method was used for antimicrobial susceptibility testing. 18 hours incubated broth culture of all the test strains were prepared and were inoculated on Muller Hinton Agar (Himedia) by making a lawn using sterile swab. Then the discs were placed over the surface of the agar and incubated at 370C for 24 hours. The zone of inhibition was measured after 24 hours of incubation. RESULTS TLC analysis of Alpinia officinarum extracts: The optimum solvent concentration was optimized, among the solvent tested Toluene: Methanol: Chloroform (1:1:1) was optimized as a better solvent combination taken to elute active compounds from the crude ethanolic extract of Alpinia officinarum. The TLC showed four fractioned spots were visualized with the Rf values 0.042, 0.286, 0.336, 0.385 (Fig –2). Antibacterial activity of ethanolic extract of Alpinia officinarum: The ethanolic extract of Alpinia officinarum showed inhibitory effect against Proteus sp. (Fig – 3), E coli (Fig – 4), Salmonella sp. (Fig – 5) and Staphylococcus aureus. (Fig – 6). The increasing zone of inhibition was identified for all the concentration of crude ethanolic extract (30µl, 60µl and 90µl). The results are shown in Table-1. DISCUSSION The rhizomes of Alpinia officinarum, is one of the major drug used in Indian systems of medicine Six diarylheptanoids, including 3 new compounds, were isolated from the rhizomes of Alpinia officinarum. The structures of the new compounds were 7-(4''-hydroxyphenyl)-1-phenyl-4-hepten-3-one, 5-methoxy-7(4''-hydroxyphenyl)-1-phenyl-3-heptanone, and 5-methoxy-1,7-diphenyl-3-heptanone detected on the basis of the spectroscopic data.12 Organic solvent extracts of Alpinia officinarum have been shown to have antimicrobial and immunomodulating properties. The rhizomes show a very good anti inflammatory effect and have carminative properties. Alpinia officinarum is used for the treatment of cold, cough, fever, inflammatory conditions, as an anticancer and anticalculi agent, as a digestive, spleen and liver tonic, and in dyspepsia, gastralgia, sea sickness and abdominal colic. Recent studies confirmed that Alpinia species possesses significant antiulcer, anticalculi, and anti-inflammatory activities.6 The ethanolic extracts of the rhizomes of Alpinia officinarum showed a significant inhibitory action against all the pathogenic organisms tested (Proteus, E coli, Klebsiella, Salmonella and Staphylococcus). This confirms the biological efficiency of crude ethanolic extracts of Alpinia officinarum and it can be further experimented and is evidence for further research. A flavonoid extracted from Alpinia officinarum shows activity against a number of gram-positive and gram-negative bacteria.8 The total extract from Alpinia officinarum rhizome showed activity against Helicobacter pylori. The total extract contains > 5% and liposol fraction contains > 30% of diphenylheptanones respectively. The extract can be used in preparation of medicaments for prophylaxis and treatment of Helicobacter pylori infection and its related diseases.9 The extracted compound galangin from Alpinia officinarum and the total flavones from the same plant show Antiviral and antitumor activity.10 Alpinia Officinanum Rhlizome extract could significantly inhibit spontaneous constriction of ileum of rabbits in vitro; exert all inhibitory effect on the contraction induced by barium chloride, histamine. It also inhibited the inflow of in cellular Ca2+ induced by ACh. This effect could be related to Ca2+ channel, M-receptor, H-receptor and direct action.11 TABLE - 1 Serial No Bacterial strains Zone of inhibition in millimeter 30µl / 3 mg 60µl / 6 mg 90µl / 9 mg 1 Proteus sp. 8 mm 10 mm 12 mm 2 Escherichia coli 7 mm 11 mm 14 mm 3 Salmonella sp. 7 mm 10 mm 12 mm 4 Staphylococcus aureus. 7 mm 12 mm 14 mm REFERENCES 1. N.Kandaswamy Pillai, History of Siddha Medicine,1-4,283,284, 1979 2. WHO Regional Consultation on Development of Traditional Medicine in the South-East Asia Region June 2005 - Document no. 16 3. ASHOK D.B.VAIDYA, Financing and Delivery of Health Care Services in India - Background Papers of the National Commission on Macroeconomics and Health, 77 4. Paul M Dewick – Medicinal Natural Products – A Biosynthetic Approach, 7,8 5. Www.Wikipedia.org 6. Journal of Medicinal and Aromatic Plant Sciences (1999), 21(4), 1155-1168. Publisher: Central Institute of Medicinal and Aromatic Plants 7. The Wealth of India – A dictionary of Indian raw materials & industrial products (CSIR) Raw materials – Vol – I:A 8. Ray, P. G.; Majumdar, S. K. Dep. Food Technol. Biochem. Eng., Jadavpur Univ., Calcutta, India. Indian Journal of Experimental Biology (1976), 14(6), 712-14. 9. Yue, Jianmin, Chengqi et al., (Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Peop. Rep. China). Faming Zhuanli Shenqing Gongkai Shuomingshu (2005), 9 pp. Written in Chinese. 10. Li, Jilai, Liu, Juyan et al., Faming Zhuanli Shenqing Gongkai Shuomingshu (2006), 14pp. 11. Tang, Chunping, Jiang et al., Zhongyao Yaoli Yu Linchuang (2006), 22(5), 23-25. Publisher: Zhongyao Yaoli Yu Linchuang Bianjibu. Journal written in Chinese. 12 . Itokawa, Hideji, Morita et al., Chemical & Pharmaceutical Bulletin (1985), 33(11), 4889-93. Journal written in English. | |||||||||||||||||||||||||||||
