Download 6. Brief resume of the intended work: 6.1: Need to the study: Alpinia

6.
Brief resume of the intended work:
6.1: Need to the study:
 Alpinia galanga, a plant in the ginger family is an herb widely used in traditional system
of medicine. Phytochemical constituents of Alpinia galanga rhizome has antitumor,
antiulcer, anticalculi, anti-microbial, anti-fungal2, antiallergic and hypoglycemic
activities3. It is reported to have gastroprotective effects8 and free radical scavenging
activity4.
 Micropropagation is a unique method for the production of uniform and disease free
plants. This technique would facilitate to obtain large number of uniform plants
irrespective of season and will serve as an alternative source of seed materials. In vitro
preservation of germplasm is also a safe method to protect the species by reducing the risk
of natural vagaries1.
 The process of micropropagation using various phytohormones in different concentration
will result in the production of large number of plantlets in short period of time
 Generic nature of plant plays a major role in defining the quantity of active principles.
Cultivation and production of secondary metabolites is constant under standard
conditions. Hence production of plant by tissue culture method will be helpful in
producing the plants of specific type of significant importance.
 Estimation of active constituent content in in vitro regenerated plant by tissue culture and
comparing it with the content of natural propogated plant has not been yet reported. Thus
the present work is an attempt to know whether the secondary cell constituent has
increased in In vitro grown plant or not.
 Considering the importance of above parameters it is concluded that research work on
tissue culture studies of Alpinia galanga plant is of significance.
6.2: Review of literature.
 Emerging buds of rhizome of Alpinia galanga produced shoots and roots simultaneously
when cultured in MS medium supplemented with Kn 3.0mgl-1. Each explanted shoot bud
produced 8 shoots in an average and roots simultaneously with in 8 weeks. Shoot
proliferation was continued even after a year by transferring each divided shoot explants
to the same media. Regenerated plants were successfully transferred to soil where they
grew well within 10-12 weeks with 80% survivality1.
 Alpinia galanga rhizome oils were screened for their anti-fungal activity against five
dermatophytes (Trichophyton mentagrophytes, T. rubrum, Microsporum canis, M. nanum
and Epidermophyton floccosum), three filamentous fungi (Aspergillus niger., A. fumigatus
and Mucor sp.) and five strains of yeasts (Saccharomyces cerevisiae, Cryptococcus
neoformans, Candida albicans, Candida tropicalis and Torulopsis glabrata). The antifungal testing was carried out by using broth microdilutions and disc gel diffusion
methods. The rhizome oil of Alpinia galanga was reported to have anti-fungal activity
against selected species of A.fumigates with a MIC value of 0.63µgμl-1. The chemical
composition of the rhizome oil was investigated by GC and GCMS methods2.
 Hypoglycemic activity was investigated in rhizome extracts of Alpinia galanga. In normal
rabbits, powdered rhizomes and its methanol, aqueous extracts significantly lowered the
blood glucose levels. In alloxan-diabetic rabbits, Alpinia galanga and its methanol,
aqueous extracts did not produced significant reduction in blood glucose levels. The
hypoglycemic effect of Alpinia galanga in normal rabbits was comparable with gliclazide.
It was concluded that Alpinia galanga produced fall in blood glucose level in normal
rabbits3.
 Methanolic extract, aqueous extract and volatile oils of the fresh rhizomes of Alpinia
galanga have been assessed for the free radical scavenging activity against 1,1-diphenyl2-picrylhydrazyl (DPPH) radical and cytotoxic activity against MCF7 (breast
adenocarcinoma) and LS174T (adenocarcinoma) cell lines. The oils of Alpinia galanga
(AGV) was analysed by GC/MS. The novel compound p-coumaryl-9-methyl ether, was
isolated from methanolic extract of Alpinia galanga4.
 Chloroform, methanol and aqueous extracts of Alpinia galanga was investigated for antibacterial activity against important pathogenic bacteria commonly associated with AIDS
infection. Inhibition growth was tested using paper disc agar diffusion method. Minimum
inhibitory concentration and Minimum bacterial concentration were determined by agar
microdilution method and agar dilution method in petridishs with millipore filters. The
gram +ve bacteria were proved to be susceptible to the chloroform extract of Alpinia
galanga and also it showed greatest inhibition zones of 29.1 and 23.7mm against S.aureus
and MRSA(Methicillin-resistant-Staphylococcus aureus). An active compound 1’acetoxy-chavicol acetate, was identified with MIC values against MRSA and
Staphylococcus aureus of 64 and 128μg/ml5.
 Four isomers of acetoxycineoles,(trans and cis)-2-3-acetoxy-1,8-cineoles, were identified
as the odorous components of the rhizomes of Alpinia galanga. Their structures were
confirmed by comparing the retention indices by GC and Mass spectra with those of
synthesized compound. The isomers presented individual odor features: the (trans and
cis)-2-isomers exhibited woody and sweet aromas. While the (trans and cis)-3-isomers
showed sweet floral and camphoraceous aromas6.
 Chemical investigation of chloroform extract of rhizomes of Alpinia galanga yielded phydroxy cinnamaldehyde and [di-(p-hydroxy-cis-styryl)]methane. The former was
isolated for the first time in nature and the latter is a new chemical component. These
compounds were characterized from spectral studies and chemical reactions7.
 The effect of 1’S-1’-acetoxy-chavicol acetate and related phenylpropanoids isolated from
rhizomes of Alpinia galanga on ethanol induced gastric lesions in rat were examined.
Among them 1’S-1’-acetoxy-chavicol acetate and 1’S-1’-acetoxyeugenol acetate
markedly inhibited the ethanol induced gastric mucosal lesions (ED50=0.61 and ca.
0.90mg/kg)8.
6.3: Main objectives of the study:
 Plant regeneration of Alpinia galanga by Tissue culture method.
 To study the effect of hormones on micropropagated plantlets.
 Study and quantify the secondary active constituent content from the in- vitro
regenerated plants and comparing it with the constituents of naturally propogated
plant.
 To study the anti-microbial and anti-fungal activity using various extracts of Alpinia
galanga rhizome.
7.
Materials and methods:
7.1: Source of the data:
The required data will be obtained from:
 Electronic data (Internet).
 Published research papers.
 Review articles from journals.
 Library of IISC, Govt. College of Pharmacy and IIHR.
7.2 Methods of collection of the data (including sampling procedure if any)
1. Collection of the plant material and authentication.
2. Data regarding the optimum requirements of hormones required for the in vitro
multiplication of plants.
3. Studies regarding the quantity of active constituent in plant regenerated through
micropropagation method and comparing it with naturally propogated plant.
4. The extracts prepared from in vitro regenerated plants are screened for its anti-microbial
and anti-fungal activities.
7.3:
Does the study require any investigations or intervention to be conducted on patients
other human or animals? If so, please describe briefly:
- No –
7.4:
Has ethical clearance been obtained from your institute in case of 7.3
-- Not applicable—
8.
List of references:
1. Borthakur M, Hazarika J, Singh RS. A protocol for micropropagation of Alpinia galanga.
Plant cell, Tissue and Org cult. 1999;55:231-233.
2. Ibrahim bin Jantan, Mohd Salleh Mohd Yassin, Chen Bee Chin, Lau Lee Chen, Ng Lee
Sim. Anti-fungal activity of the Essential Oils of Nine Zingiberaceae Species.
Pharmaceutical Biology. 2003;41(5):392-397.
3. Akhtar MS, Khan MA, Malik MT. Hypoglycaemic activity of Alpinia galanga rhizome
and its extracts in rabbits. Fitoterapia. 2002;73:623-628.
4. Sariga Zaeoung, Anuchit Plubrukarn, Niwat Keawpradub. Cytotoxic and free radical
scavenging activities of Zingiberaceous rhizomes. J Sci. Technol. 2005;27(4):799-812.
5. Supayang piyawan Voravuthikunchai, Souwalak Phongpaichit, Sanan Subhadhirasakul.
Evaluation of Anti-bacterial Activities of Medicinal plants widely used among AIDS
Patients in Thailand. Pharmaceutical Biology. 2005;43(8):701-706.
6. Kikue Kubota, Kae Nakamura (Murayama), Akio Kobayashi. Acetoxy-1,8-cineoles as
Aroma Constituents of Alpinia galanga Willd. J Agric. Food Chem 1998;46(12):52445247.
7. Barik BR, Kundu AB, Dey AK. Two phenolic constituents from Alpinia galanga
rhizomes. Phytochemistry. 1987;26(7):2126-2127.
8. Hisashi Matsuda, Yutana Pongpiriyadacha, Toshio Morikawa, MomotaroOchi ,Masayuki
Yoshikawa. Gastroprotective effects of Phenylpropanoids from the rhizomes of Alpinia
galanga in rats: structural requirements and mode of action. European Journal of
Pharmacology. 2003;471(1):59-67.